LRRK2 Antibody - BSA Free

Western Blot: LRRK2 AntibodyBSA Free [NB300-268]

Western Blot: LRRK2 Antibody [NB300-268] - LRRK2 N-terminal sequences show increased aggregation. Western blots of cell lysates of SH-SY5Y cells transfected with indicated LRRK2 construct showed equal expression of the LRRK2 protein with anti-HA (top panel) or anti-C-terminal- (bottom panel) LRRK2 antibody. Equal lysate loading is indicated by Beta-actin loading control. Image collected and cropped by CiteAb from the following publication (//doi.org/10.1371/journal.pone.0045149) licensed under a CC-BY license.

Immunohistochemistry-Paraffin: LRRK2 Antibody - BSA Free [NB300-268]

Immunohistochemistry-Paraffin: LRRK2 Antibody [NB300-268] - Specificity of immunostaining for LRRK2. Specificity of the antibodies used for immunocytochemistry was determined by performing adsorptions with the corresponding peptide sequences. Immunochemical localization of LBs (arrows) by Ab1 against LRRK2900-100 (left) was blocked using respective peptide antigens (right). Adjacent sections with LB within pigmented neurons from the substantia nigra of a case of PD were used. Scale bar: 20 um. Image collected and cropped by CiteAb from the following publication (https://molecularneurodegeneration.biomedcentral.com/articles/10.1186/1750-1326-1-17), licensed under a CC-BY license.

Immunohistochemistry-Paraffin: LRRK2 Antibody - BSA Free [NB300-268]

Immunohistochemistry-Paraffin: LRRK2 Antibody [NB300-268] - Immunocytochemistry of LRRK2 in PD using antibodies (Ab1, Ab4) raised against sequences corresponding to various regions shown on the schematic diagram of LRRK2 (red bars) were used on brainstem sections of PD. Scale bars: 10 um (top); E-L = 100 um (bottom). Image collected and cropped by CiteAb from the following publication (https://molecularneurodegeneration.biomedcentral.com/articles/10.1186/1750-1326-1-17), licensed under a CC-BY license.

Flow Cytometry: LRRK2 Antibody - BSA Free [NB300-268]

Flow Cytometry: LRRK2 Antibody [NB300-268] - Analysis using the Alexa Fluor 647 conjugate of NB300-268. Staining of LRRK2 in normal human peripheral blood cells (anticoagulated Lithium-Heparin) using anti-LRRK2 antibody conjugated with AF647. The primary antibody was used at a dilution of 1:200 and incubated for 20 min at room temperature. Image from verified customer review.

Immunohistochemistry-Paraffin: LRRK2 Antibody - BSA Free [NB300-268]

Immunohistochemistry-Paraffin: LRRK2 Antibody [NB300-268] - LRRK2 accumulates in globules in alphaS tg mice. Triple immunofluorescence for alphaS, LRRK2 and Rab5B for basal ganglia in alphaS tg mice. LRRK2 and Rab5B were colocalized in axon terminal (arrow), but were not colocalized in the alphaS-globule (arrowhead) Scale bar = 10 um for all panels. Image collected and cropped by CiteAb from the following publication (https://molecularbrain.biomedcentral.com/articles/10.1186/1756-6606-5-34), licensed under a CC-BY license.

Immunocytochemistry/ Immunofluorescence: LRRK2 Antibody - BSA Free [NB300-268]

Immunocytochemistry/Immunofluorescence: LRRK2 Antibody [NB300-268] - HeLa cells were fixed in 4% paraformaldehyde for 10 minutes and permeabilized in 0.05% Triton X-100 in PBS for 5 minutes. The cells were incubated with anti-LRRK2 Antibody NB300-268 at 2 ug/ml overnight at 4C and detected with an anti-mouse Dylight 488 (Green) at a 1:1000 dilution for 60 minutes. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 100X objective and digitally deconvolved.

Flow Cytometry: LRRK2 Antibody - BSA Free [NB300-268]

Flow Cytometry: LRRK2 Antibody [NB300-268] - An intracellular stain was performed on U2OS cells with LRRK2 Antibody NB300-268 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (SA5-10033, Thermo Fisher).

Immunocytochemistry/ Immunofluorescence: LRRK2 Antibody - BSA Free [NB300-268]

Immunocytochemistry/Immunofluorescence: LRRK2 Antibody [NB300-268] - LRRK2 antibody was tested in SH-SY-5Y cells at a 1:200 dilution against DyLight 488 (Green). Alpha tubulin and nuclei were counterstained against DyLight 568 (Red), and DAPI (Blue), respectively.

Immunohistochemistry-Paraffin: LRRK2 Antibody - BSA Free [NB300-268]

Immunohistochemistry-Paraffin: LRRK2 Antibody [NB300-268] - Staining of human brain, Putamen, Neurons and Glia using a 60X magnification.

Flow Cytometry: LRRK2 Antibody - BSA Free [NB300-268]

Flow Cytometry: LRRK2 Antibody [NB300-268] - An intracellular stain was performed on U2OS cells with LRRK Antibody NB300-268AF647 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.

Flow Cytometry: LRRK2 Antibody - BSA Free [NB300-268]

Flow Cytometry: LRRK2 Antibody [NB300-268] - An intracellular stain was performed on Neuro2a cells with LRRK2 Antibody NB300-268 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeabilized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes at room temperature, followed by Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550 (SA5-10033, Thermo Fisher).

Western Blot: LRRK2 Antibody - BSA Free [NB300-268] -

LRRK2 kinase inhibition blocks Tat-induced S935 phosphorylation and inflammatory cytokine expression in BV-2 cells. (A) Western blot depicting pS935-LRRK2 and total LRRK2 protein levels 12 hours after saline or Tat-treatment ± LRRK2 kinase inhibition (LRRK2i).

Immunocytochemistry/Immunofluorescence: LRRK2 Antibody - BSA Free [NB300-268] -

The interaction and phosphorylation of LRRK2 and LC1. (A) Cellular co-localisation of endogenous LRRK2 and MAP1B (LC1). SKNSH cells were labelled with LRRK2 (green) and LC1 (red) antibodies and signals were detected by immunofluorescence. Detailed figure in Additional file 1.

Western Blot: LRRK2 Antibody - BSA Free [NB300-268] -

Western Blot: LRRK2 Antibody - BSA Free [NB300-268] - Western blotting of LRRK2. A. Recombinant LRRK2 (arrowhead) from transfected (+) HEK 293T & M17 cells was specifically recognized by all four LRRK2 antibodies (Ab1, Ab2, Ab3, & Ab4) used in this study. LRRK2 was not recognized in non-transfected cells (-). B. Recombinant LRRK2 (arrowhead) from transfected M17 cells is detected by anti-LRRK2 Ab4 (lane 1), but not after absorption of the antibody with its peptide antigen (lane 2). Brain LRRK2 was recognized by anti-LRRK2 Ab4 in two controls (lanes 3 & 4) & two PD cases (lanes 5 & 6). Cell lysates (10 μg protein) & brain homogenates (100 μg protein) were prepared & loaded on 6% SDS-PAGE gels for Western blot analysis using anti-LRRK2 antibodies Ab1–Ab4 in (A) & Ab4 in (B). Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/17137507), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Flow Cytometry: LRRK2 Antibody - BSA Free [NB300-268] -

Flow Cytometry: LRRK2 Antibody - BSA Free [NB300-268] - Deletion of LRRK2 attenuates Mn-induced apoptosis & cell death in RAW 264.7 cells.(A) LRRK2 WT or KO RAW 264.7 cells were treated with Mn (250 μM) for the designated times, followed by flow cytometry analysis to determine Mn-induced apoptosis. Both early & late apoptotic cells (Q2 & Q3) were measured. (B) At the end of Mn exposure, cell viability was determined by MTT assay. @@@, p < 0.001; ###, p < 0.001; ***, p < 0.001 compared to the control (one-way ANOVA followed by Tukey’s post hoc test; n = 3, for apoptosis assay; n = 6, for MTT assay). The data shown are representative of 3 independent experiments. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30645642), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: LRRK2 Antibody - BSA Free [NB300-268] -

Immunocytochemistry/ Immunofluorescence: LRRK2 Antibody - BSA Free [NB300-268] - LRRK2 accumulates in globules in alphaS tg mice. (a & b) Double immunofluorescence for alphaS with parkin, PINK1, DJ-1, LRRK2, or negative control (the immunopeptide-preabsorbed anti-LRRK2 antibody) in alphaS tg mice (a) & P123H betaS tg mice (b). Note that alphaS-globules were immunopositive for LRRK2 (~79%, n = 22), whereas P123H betaS globules were negative for LRRK2. Representative images are shown for the thalamus ( alphaS) & basal ganglia (P123H betaS). Scale bar = 5 μm for all panels. (c) Triple immunofluorescence for alphaS, LRRK2 & Rab5B for basal ganglia in alphaS tg mice. LRRK2 & Rab5B were colocalized in axon terminal (arrow), but were not colocalized in the alphaS-globule (arrowhead) Scale bar = 10 μm for all panels. Image collected & cropped by CiteAb from the following publication (https://molecularbrain.biomedcentral.com/articles/10.1186/1756-6606-5-34), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry-Paraffin: LRRK2 Antibody - BSA Free [NB300-268] -

Immunohistochemistry-Paraffin: LRRK2 Antibody - BSA Free [NB300-268] - Specificity of immunostaining for LRRK2. Specificity of the antibodies used for immunocytochemistry was determined by performing adsorptions with the corresponding peptide sequences. Immunochemical localization of LBs (arrows) by Ab4 against LRRK22500–2527 (A) & Ab1 against LRRK2900–100 (C) was blocked using respective peptide antigens (B & D). Adjacent sections with LB within pigmented neurons from the substantia nigra of a case of PD were used. Scale bar: 20 μm. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/17137507), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry-Paraffin: LRRK2 Antibody - BSA Free [NB300-268] -

Immunohistochemistry-Paraffin: LRRK2 Antibody - BSA Free [NB300-268] - Immunocytochemistry of LRRK2 in DLB. Cortical LBs (arrows) in DLB were positive for LRRK2 using both Ab4 against LRRK22500–2527 (A), & Ab1 against LRRK2900–100 (B). Neuronal cytoplasm (C) was also strongly labelled throughout the cortex by Ab4 & with a consistently more granular pattern by Ab1 (inset in C) in cases of DLB. Control cases display similar, though less intense neuronal labeling (D). Biochemically purified cortical LBs were strongly positive for staining by Ab4 (E) as well as by anti-alpha -synuclein (F), while they were unstained when omitting primary antibody (G). Large vessels (H) were also intensely labelled by Ab4. Scale bars: A, B = 25 μm; C, D = 100 μm; E-G = 25 μm; H = 50 μm. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/17137507), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry-Paraffin: LRRK2 Antibody - BSA Free [NB300-268] -

Immunohistochemistry-Paraffin: LRRK2 Antibody - BSA Free [NB300-268] - Immunocytochemistry of LRRK2 in DLB. Cortical LBs (arrows) in DLB were positive for LRRK2 using both Ab4 against LRRK22500–2527 (A), & Ab1 against LRRK2900–100 (B). Neuronal cytoplasm (C) was also strongly labelled throughout the cortex by Ab4 & with a consistently more granular pattern by Ab1 (inset in C) in cases of DLB. Control cases display similar, though less intense neuronal labeling (D). Biochemically purified cortical LBs were strongly positive for staining by Ab4 (E) as well as by anti-alpha -synuclein (F), while they were unstained when omitting primary antibody (G). Large vessels (H) were also intensely labelled by Ab4. Scale bars: A, B = 25 μm; C, D = 100 μm; E-G = 25 μm; H = 50 μm. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/17137507), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: LRRK2 Antibody - BSA Free [NB300-268] -

Immunocytochemistry/ Immunofluorescence: LRRK2 Antibody - BSA Free [NB300-268] - LRRK2 accumulates in globules in alphaS tg mice. (a & b) Double immunofluorescence for alphaS with parkin, PINK1, DJ-1, LRRK2, or negative control (the immunopeptide-preabsorbed anti-LRRK2 antibody) in alphaS tg mice (a) & P123H betaS tg mice (b). Note that alphaS-globules were immunopositive for LRRK2 (~79%, n = 22), whereas P123H betaS globules were negative for LRRK2. Representative images are shown for the thalamus ( alphaS) & basal ganglia (P123H betaS). Scale bar = 5 μm for all panels. (c) Triple immunofluorescence for alphaS, LRRK2 & Rab5B for basal ganglia in alphaS tg mice. LRRK2 & Rab5B were colocalized in axon terminal (arrow), but were not colocalized in the alphaS-globule (arrowhead) Scale bar = 10 μm for all panels. Image collected & cropped by CiteAb from the following publication (https://molecularbrain.biomedcentral.com/articles/10.1186/1756-6606-5-34), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: LRRK2 Antibody - BSA Free [NB300-268] -

Immunocytochemistry/ Immunofluorescence: LRRK2 Antibody - BSA Free [NB300-268] - LRRK2 accumulates in globules in alphaS tg mice. (a & b) Double immunofluorescence for alphaS with parkin, PINK1, DJ-1, LRRK2, or negative control (the immunopeptide-preabsorbed anti-LRRK2 antibody) in alphaS tg mice (a) & P123H betaS tg mice (b). Note that alphaS-globules were immunopositive for LRRK2 (~79%, n = 22), whereas P123H betaS globules were negative for LRRK2. Representative images are shown for the thalamus ( alphaS) & basal ganglia (P123H betaS). Scale bar = 5 μm for all panels. (c) Triple immunofluorescence for alphaS, LRRK2 & Rab5B for basal ganglia in alphaS tg mice. LRRK2 & Rab5B were colocalized in axon terminal (arrow), but were not colocalized in the alphaS-globule (arrowhead) Scale bar = 10 μm for all panels. Image collected & cropped by CiteAb from the following publication (https://molecularbrain.biomedcentral.com/articles/10.1186/1756-6606-5-34), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LRRK2 Antibody - BSA Free [NB300-268] -

Western Blot: LRRK2 Antibody - BSA Free [NB300-268] - Western blotting of LRRK2. A. Recombinant LRRK2 (arrowhead) from transfected (+) HEK 293T & M17 cells was specifically recognized by all four LRRK2 antibodies (Ab1, Ab2, Ab3, & Ab4) used in this study. LRRK2 was not recognized in non-transfected cells (-). B. Recombinant LRRK2 (arrowhead) from transfected M17 cells is detected by anti-LRRK2 Ab4 (lane 1), but not after absorption of the antibody with its peptide antigen (lane 2). Brain LRRK2 was recognized by anti-LRRK2 Ab4 in two controls (lanes 3 & 4) & two PD cases (lanes 5 & 6). Cell lysates (10 μg protein) & brain homogenates (100 μg protein) were prepared & loaded on 6% SDS-PAGE gels for Western blot analysis using anti-LRRK2 antibodies Ab1–Ab4 in (A) & Ab4 in (B). Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/17137507), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry-Paraffin: LRRK2 Antibody - BSA Free [NB300-268] -

Immunohistochemistry-Paraffin: LRRK2 Antibody - BSA Free [NB300-268] - Immunocytochemistry of LRRK2 in DLB. Cortical LBs (arrows) in DLB were positive for LRRK2 using both Ab4 against LRRK22500–2527 (A), & Ab1 against LRRK2900–100 (B). Neuronal cytoplasm (C) was also strongly labelled throughout the cortex by Ab4 & with a consistently more granular pattern by Ab1 (inset in C) in cases of DLB. Control cases display similar, though less intense neuronal labeling (D). Biochemically purified cortical LBs were strongly positive for staining by Ab4 (E) as well as by anti-alpha -synuclein (F), while they were unstained when omitting primary antibody (G). Large vessels (H) were also intensely labelled by Ab4. Scale bars: A, B = 25 μm; C, D = 100 μm; E-G = 25 μm; H = 50 μm. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/17137507), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry-Paraffin: LRRK2 Antibody - BSA Free [NB300-268] -

Immunohistochemistry-Paraffin: LRRK2 Antibody - BSA Free [NB300-268] - Immunocytochemistry of LRRK2 in DLB. Cortical LBs (arrows) in DLB were positive for LRRK2 using both Ab4 against LRRK22500–2527 (A), & Ab1 against LRRK2900–100 (B). Neuronal cytoplasm (C) was also strongly labelled throughout the cortex by Ab4 & with a consistently more granular pattern by Ab1 (inset in C) in cases of DLB. Control cases display similar, though less intense neuronal labeling (D). Biochemically purified cortical LBs were strongly positive for staining by Ab4 (E) as well as by anti-alpha -synuclein (F), while they were unstained when omitting primary antibody (G). Large vessels (H) were also intensely labelled by Ab4. Scale bars: A, B = 25 μm; C, D = 100 μm; E-G = 25 μm; H = 50 μm. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/17137507), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry-Paraffin: LRRK2 Antibody - BSA Free [NB300-268] -

Immunohistochemistry-Paraffin: LRRK2 Antibody - BSA Free [NB300-268] - Immunocytochemistry of LRRK2 in DLB. Cortical LBs (arrows) in DLB were positive for LRRK2 using both Ab4 against LRRK22500–2527 (A), & Ab1 against LRRK2900–100 (B). Neuronal cytoplasm (C) was also strongly labelled throughout the cortex by Ab4 & with a consistently more granular pattern by Ab1 (inset in C) in cases of DLB. Control cases display similar, though less intense neuronal labeling (D). Biochemically purified cortical LBs were strongly positive for staining by Ab4 (E) as well as by anti-alpha -synuclein (F), while they were unstained when omitting primary antibody (G). Large vessels (H) were also intensely labelled by Ab4. Scale bars: A, B = 25 μm; C, D = 100 μm; E-G = 25 μm; H = 50 μm. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/17137507), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LRRK2 Antibody - BSA Free [NB300-268] -

Western Blot: LRRK2 Antibody - BSA Free [NB300-268] - LRRK2 deletion attenuates Mn-induced pro-apoptotic protein levels.(A) After cells (LRRK2 WT & LRRK2 KO) were exposed to Mn (250 μM) for up to 24 h, total protein was extracted & followed by western blot analysis to determine protein levels of Bax & Daxx. beta-actin was used as a loading control. (B) The expression levels of Bax & Daxx modulated by Mn were quantified relative to its control levels in LRRK2 WT & KO cells. **, p < 0.01; ***, p < 0.001 (one-way ANOVA followed by Tukey’s post hoc test; n = 3). The data shown are representative of 3 independent experiments. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30645642), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: LRRK2 Antibody - BSA Free [NB300-268] -

Immunocytochemistry/ Immunofluorescence: LRRK2 Antibody - BSA Free [NB300-268] - The interaction & phosphorylation of LRRK2 & LC1. (A) Cellular co-localisation of endogenous LRRK2 & MAP1B (LC1). SKNSH cells were labelled with LRRK2 (green) & LC1 (red) antibodies & signals were detected by immunofluorescence. Detailed figure in Additional file 1. (B) LRRK2 kinase assay. The LRRK2 wild-typed (WT) kinase interacted & phosphorylated the LC1 domain unlike the LRRK2 kinase-dead protein. (C) Western blotting of LRRK2 & LC1 proteins after a kinase assay. Both LRRK2 & LC1 proteins were probed for phospho-threonine & phospho-serine signals respectively. Image collected & cropped by CiteAb from the following publication (http://molecularbrain.biomedcentral.com/articles/10.1186/1756-6606-7-29), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LRRK2 Antibody - BSA Free [NB300-268] -

Western Blot: LRRK2 Antibody - BSA Free [NB300-268] - Mn increases LRRK2 kinase activity & expression in RAW 264.7 & HMC3 cells.(A) Two-hundred (200) μg of protein were collected from cell lysates, followed by western blot analysis to determine the presence of LRRK2 in LRRK2 WT, KO RAW 264.7 & HMC3 cells. beta-actin was used as a loading control. (B,C) LRRK2 WT or HMC3 cells were treated with Mn (250 μM) for the designated times, followed by protein extraction & western blot analysis as described in the Methods section. Protein levels of LRRK2 & phosphorylated LRRK2 (S1292) in LRRK2 WT RAW 264.7 (B) & HMC3 (C) cells were quantified. (D,E) Effect of Mn on LRRK2 mRNA levels in LRRK2 WT RAW 264.7 (D) & HMC3 (E) cells were assessed as described in the Methods section. GAPDH was used as a loading control. (F) After pre-treatment with LRRK2 inhibitors GSK (1 μM) & MLi-2 (50 nM) for 90 min, RAW 264.7 cells were exposed to Mn (250 μM) for 20 min, followed by western blot analysis to detect phosphorylation of LRRK2 (S1292). ###, p < 0.001; *, p < 0.05; **, p < 0.01; ***, p < 0.001 compared to the control (one-way ANOVA followed by Tukey’s post hoc test; n = 3). The data shown are representative of 3 independent experiments. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30645642), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry-Paraffin: LRRK2 Antibody - BSA Free [NB300-268] -

Immunohistochemistry-Paraffin: LRRK2 Antibody - BSA Free [NB300-268] - Immunocytochemistry of LRRK2 in PD. Four antibodies raised against sequences corresponding to various regions shown on the schematic diagram of LRRK2 (red bars) were used on brainstem sections of PD (A-H) & age-matched controls (I-L). Intense immunolabeling of brainstem LBs in cases of PD was seen with Ab1 against LRRK2900–100 (A) & Ab4 against LRRK22500–2527(D). Both rim (A & D) & core (inset in D) of LBs were recognized. In contrast, LBs were not labelled in any case using antibodies directed against LRRK21246–1265 (Ab2, B) or LRRK21838–2133 (Ab3, C), for which the antigenic sites are located within the folded domains. The cell bodies of both pigmented & no-pigmented neurons (arrows) as well as axons (arrowheads) contain LRRK2, seen only using antibodies against sites outside the folded domains (Ab1, E & Ab4, H). In contrast, Ab4 staining was much less intense in control tissue (L). The muscle layer of both large & small vessels was consistently found to contain high levels of LRRK2 in almost all PD cases (seen in E, G, & H) & strikingly the vessels are the only structure immunolabeled with Ab3 recognizing LRRK21838–2133 in all cases studied (G & K). Ab2 to LRRK21246–1265 did not recognize LBs, cell bodies or vessels in any case (B, F, & J). Scale bars: A-D & inset = 10 μm; E-L = 100 μm. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/17137507), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry-Paraffin: LRRK2 Antibody - BSA Free [NB300-268] -

Immunohistochemistry-Paraffin: LRRK2 Antibody - BSA Free [NB300-268] - Immunocytochemistry of LRRK2 in PD. Four antibodies raised against sequences corresponding to various regions shown on the schematic diagram of LRRK2 (red bars) were used on brainstem sections of PD (A-H) & age-matched controls (I-L). Intense immunolabeling of brainstem LBs in cases of PD was seen with Ab1 against LRRK2900–100 (A) & Ab4 against LRRK22500–2527(D). Both rim (A & D) & core (inset in D) of LBs were recognized. In contrast, LBs were not labelled in any case using antibodies directed against LRRK21246–1265 (Ab2, B) or LRRK21838–2133 (Ab3, C), for which the antigenic sites are located within the folded domains. The cell bodies of both pigmented & no-pigmented neurons (arrows) as well as axons (arrowheads) contain LRRK2, seen only using antibodies against sites outside the folded domains (Ab1, E & Ab4, H). In contrast, Ab4 staining was much less intense in control tissue (L). The muscle layer of both large & small vessels was consistently found to contain high levels of LRRK2 in almost all PD cases (seen in E, G, & H) & strikingly the vessels are the only structure immunolabeled with Ab3 recognizing LRRK21838–2133 in all cases studied (G & K). Ab2 to LRRK21246–1265 did not recognize LBs, cell bodies or vessels in any case (B, F, & J). Scale bars: A-D & inset = 10 μm; E-L = 100 μm. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/17137507), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LRRK2 Antibody - BSA Free [NB300-268] -

Western Blot: LRRK2 Antibody - BSA Free [NB300-268] - Mn increases LRRK2 kinase activity & expression in RAW 264.7 & HMC3 cells.(A) Two-hundred (200) μg of protein were collected from cell lysates, followed by western blot analysis to determine the presence of LRRK2 in LRRK2 WT, KO RAW 264.7 & HMC3 cells. beta-actin was used as a loading control. (B,C) LRRK2 WT or HMC3 cells were treated with Mn (250 μM) for the designated times, followed by protein extraction & western blot analysis as described in the Methods section. Protein levels of LRRK2 & phosphorylated LRRK2 (S1292) in LRRK2 WT RAW 264.7 (B) & HMC3 (C) cells were quantified. (D,E) Effect of Mn on LRRK2 mRNA levels in LRRK2 WT RAW 264.7 (D) & HMC3 (E) cells were assessed as described in the Methods section. GAPDH was used as a loading control. (F) After pre-treatment with LRRK2 inhibitors GSK (1 μM) & MLi-2 (50 nM) for 90 min, RAW 264.7 cells were exposed to Mn (250 μM) for 20 min, followed by western blot analysis to detect phosphorylation of LRRK2 (S1292). ###, p < 0.001; *, p < 0.05; **, p < 0.01; ***, p < 0.001 compared to the control (one-way ANOVA followed by Tukey’s post hoc test; n = 3). The data shown are representative of 3 independent experiments. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30645642), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LRRK2 Antibody - BSA Free [NB300-268] -

Western Blot: LRRK2 Antibody - BSA Free [NB300-268] - Mn increases LRRK2 kinase activity & expression in RAW 264.7 & HMC3 cells.(A) Two-hundred (200) μg of protein were collected from cell lysates, followed by western blot analysis to determine the presence of LRRK2 in LRRK2 WT, KO RAW 264.7 & HMC3 cells. beta-actin was used as a loading control. (B,C) LRRK2 WT or HMC3 cells were treated with Mn (250 μM) for the designated times, followed by protein extraction & western blot analysis as described in the Methods section. Protein levels of LRRK2 & phosphorylated LRRK2 (S1292) in LRRK2 WT RAW 264.7 (B) & HMC3 (C) cells were quantified. (D,E) Effect of Mn on LRRK2 mRNA levels in LRRK2 WT RAW 264.7 (D) & HMC3 (E) cells were assessed as described in the Methods section. GAPDH was used as a loading control. (F) After pre-treatment with LRRK2 inhibitors GSK (1 μM) & MLi-2 (50 nM) for 90 min, RAW 264.7 cells were exposed to Mn (250 μM) for 20 min, followed by western blot analysis to detect phosphorylation of LRRK2 (S1292). ###, p < 0.001; *, p < 0.05; **, p < 0.01; ***, p < 0.001 compared to the control (one-way ANOVA followed by Tukey’s post hoc test; n = 3). The data shown are representative of 3 independent experiments. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30645642), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LRRK2 Antibody - BSA Free [NB300-268] -

Western Blot: LRRK2 Antibody - BSA Free [NB300-268] - Mn increases LRRK2 kinase activity & expression in RAW 264.7 & HMC3 cells.(A) Two-hundred (200) μg of protein were collected from cell lysates, followed by western blot analysis to determine the presence of LRRK2 in LRRK2 WT, KO RAW 264.7 & HMC3 cells. beta-actin was used as a loading control. (B,C) LRRK2 WT or HMC3 cells were treated with Mn (250 μM) for the designated times, followed by protein extraction & western blot analysis as described in the Methods section. Protein levels of LRRK2 & phosphorylated LRRK2 (S1292) in LRRK2 WT RAW 264.7 (B) & HMC3 (C) cells were quantified. (D,E) Effect of Mn on LRRK2 mRNA levels in LRRK2 WT RAW 264.7 (D) & HMC3 (E) cells were assessed as described in the Methods section. GAPDH was used as a loading control. (F) After pre-treatment with LRRK2 inhibitors GSK (1 μM) & MLi-2 (50 nM) for 90 min, RAW 264.7 cells were exposed to Mn (250 μM) for 20 min, followed by western blot analysis to detect phosphorylation of LRRK2 (S1292). ###, p < 0.001; *, p < 0.05; **, p < 0.01; ***, p < 0.001 compared to the control (one-way ANOVA followed by Tukey’s post hoc test; n = 3). The data shown are representative of 3 independent experiments. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30645642), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: LRRK2 Antibody - BSA Free [NB300-268] -

Western Blot: LRRK2 Antibody - BSA Free [NB300-268] - LRRK2 kinase inhibition blocks Tat-induced S935 phosphorylation & inflammatory cytokine expression in BV-2 cells. (A) Western blot depicting pS935-LRRK2 & total LRRK2 protein levels 12 hours after saline or Tat-treatment ± LRRK2 kinase inhibition (LRRK2i). (B) Tat treatment significantly increased pS935-LRRK2, which was attenuated by LRRK2i. (C) We measured TNF-alpha, IL-6, MCP-1 & IL-10 mRNA levels by qRT-PCR 12 hours post saline or Tat treatment ± LRRK2i. LRRK2i attenuated Tat-induced TNF-alpha, IL-6, MCP-1 & IL-10 expression for all groups (*P <0.05, ***P <0.001, one way ANOVA, Newman-Keuls post-test). ANOVA, analysis of variance; LRRK2, leucine-rich repeat kinase2; pS935, phosphorylation of serine 935; Tat, trans activator of transcription. Image collected & cropped by CiteAb from the following publication (http://jneuroinflammation.biomedcentral.com/articles/10.1186/1742-2094-9-261), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunohistochemistry-Paraffin: LRRK2 Antibody - BSA Free [NB300-268] -

Immunohistochemistry-Paraffin: LRRK2 Antibody - BSA Free [NB300-268] - Immunocytochemistry of LRRK2 in PD. Four antibodies raised against sequences corresponding to various regions shown on the schematic diagram of LRRK2 (red bars) were used on brainstem sections of PD (A-H) & age-matched controls (I-L). Intense immunolabeling of brainstem LBs in cases of PD was seen with Ab1 against LRRK2900–100 (A) & Ab4 against LRRK22500–2527(D). Both rim (A & D) & core (inset in D) of LBs were recognized. In contrast, LBs were not labelled in any case using antibodies directed against LRRK21246–1265 (Ab2, B) or LRRK21838–2133 (Ab3, C), for which the antigenic sites are located within the folded domains. The cell bodies of both pigmented & no-pigmented neurons (arrows) as well as axons (arrowheads) contain LRRK2, seen only using antibodies against sites outside the folded domains (Ab1, E & Ab4, H). In contrast, Ab4 staining was much less intense in control tissue (L). The muscle layer of both large & small vessels was consistently found to contain high levels of LRRK2 in almost all PD cases (seen in E, G, & H) & strikingly the vessels are the only structure immunolabeled with Ab3 recognizing LRRK21838–2133 in all cases studied (G & K). Ab2 to LRRK21246–1265 did not recognize LBs, cell bodies or vessels in any case (B, F, & J). Scale bars: A-D & inset = 10 μm; E-L = 100 μm. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/17137507), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Flow Cytometry: LRRK2 Antibody - BSA Free [NB300-268] -

Flow Cytometry: LRRK2 Antibody - BSA Free [NB300-268] - Inhibition of LRRK2 kinase activity attenuates Mn-induced apoptosis.(A) After pre-treatment with GSK (1 μM) for 90 min, cells (HMC3) were exposed to Mn (250 μM) for the designated time periods, followed by annexin V & PI staining & flow cytometry analysis to determine apoptosis. Early & late apoptotic cells (Q2 & Q3) were analyzed. (B,C) After pre-treatment with GSK (1 μM) for 90 min, cells (LRRK2 WT RAW 264.7 & HMC3) were exposed to Mn for designated time periods, followed by the MTT assay to determine cell viability, as described in the Methods section, (@@@, p < 0.001; *, p < 0.05; ***, p < 0.001 compared to the control (one-way ANOVA followed by Tukey’s post hoc test; n = 3, apoptosis assay; n = 6, MTT assay). The data shown are representative of 3 independent experiments. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30645642), licensed under a CC-BY license. Not internally tested by Novus Biologicals.