Actin Antibody (mAbGEa)

Immunohistochemistry-Paraffin: Actin Antibody (mAbGEa) [NB100-74340]

Immunohistochemistry-Paraffin: Actin Antibody (mAbGEa) [NB100-74340] - Both normal and cancer biopsies of deparaffinized Human tonsil tissues.

Flow Cytometry: Actin Antibody (mAbGEa) [NB100-74340]

Flow Cytometry: Actin Antibody (mAbGEa) [NB100-74340] - Analysis of HeLa cells using mouse Actin antibody (Orange) and Isotype control Antibody (Blue).

Western Blot: Actin Antibody (mAbGEa) [NB100-74340]

Western Blot: Actin Antibody (mAbGEa) [NB100-74340] - Increased relative abundance of Park7 in Lp(a) mice. Western blots were used to validate the comparative proteomic analysis between the Lp(a) and wildtype mice for the Park7 protein. Pooled liver protein extracts (n = 4 livers per pool) were separated by SDS PAGE in multiple replicates (n = 7) and transferred onto nitrocellulose membrane. Membranes were probed with an anti-Park7 antibody using an anti-actin antibody as a loading control. Liver protein extracts were the same as those used for Figure 4(a) as well as fresh liver protein extracts from new mice of the same age, sex, and genotype. Image collected and cropped by CiteAb from the following publication (null), licensed under a CC-BY license.

Immunocytochemistry/ Immunofluorescence: Actin Antibody (mAbGEa) [NB100-74340]

Immunocytochemistry/Immunofluorescence: Actin Antibody (mAbGEa) [NB100-74340] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.05% Triton-X100. The cells were incubated with anti-Actin Antibody (mAbGEa) at 2 ug/ml overnight at 4C and detected with an anti-mouse Dylight 488 (Green) at a 1:500 dilution. Actin was detected with Phalloidin 568 (Red) at a 1:200 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Flow (Intracellular): Actin Antibody (mAbGEa) [NB100-74340]

Flow (Intracellular): Actin Antibody (mAbGEa) [NB100-74340] - An intracellular stain was performed on A549 cells with NB100-74340 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes at room temperature, followed by mouse IgM Alexa Fluor 488-conjugated secondary antibody.

Western Blot: Actin Antibody (mAbGEa) [NB100-74340]

Western Blot: Actin Antibody (mAbGEa) [NB100-74340] - Analysis of Actin expression in 2) HeLa, 3) NTERA-2, 4) A431, 5) HepG2, 6) MCF7, 7) NIH-3T3, 8) PC-12 and 9) COS-7 whole cell lysates.

Western Blot: Actin Antibody (mAbGEa) [NB100-74340]

Western Blot: Actin Antibody (mAbGEa) [NB100-74340] - Analysis of Jurkat and CHO cell lysates using actin antibody [NB100-74340] at 1:100.

Immunocytochemistry/ Immunofluorescence: Actin Antibody (mAbGEa) [NB100-74340]

Immunocytochemistry/Immunofluorescence: Actin Antibody (mAbGEa) [NB100-74340] - Actin was detected in NIH-3T3 cells fixed with methanol using mouse anti-mouse beta-Actin monoclonal antibody (NB100-74340) at 1:1800. Cells were stained using Northern Lights 557 conjugated anti-mouse secondary antibody (NL007) and counterstained with DAPI.

Immunocytochemistry/ Immunofluorescence: Actin Antibody (mAbGEa) [NB100-74340]

Immunocytochemistry/Immunofluorescence: Actin Antibody (mAbGEa) [NB100-74340] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.05% Triton X-100. The cells were incubated with anti-Actin (mAbGEa) at 2 ug/mL overnight at 4C and detected with an anti-mouse IgM DyLight 488 (Green) at 1:500. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Immunohistochemistry-Paraffin: Actin Antibody (mAbGEa) [NB100-74340]

Immunohistochemistry-Paraffin: Actin Antibody (mAbGEa) [NB100-74340] - Analysis of Actin on human breast cancer tissue using DAB with hematoxylin counterstain.

Immunohistochemistry-Paraffin: Actin Antibody (mAbGEa) [NB100-74340]

Immunohistochemistry-Paraffin: Actin Antibody (mAbGEa) [NB100-74340] - Both normal and cancer biopsies of deparaffinized Human colon carcinoma tissues.

Immunohistochemistry-Paraffin: Actin Antibody (mAbGEa) [NB100-74340]

Immunohistochemistry-Paraffin: Actin Antibody (mAbGEa) [NB100-74340] - Both normal and cancer biopsies of deparaffinized Human skeletal muscle tissues.

Western Blot: Actin Antibody (mAbGEa) [NB100-74340] -

Western Blot: Actin Antibody (mAbGEa) [NB100-74340] - Lp(a) upregulates GPx1 & Prdx6 expression in human HepG2 cells. HepG2 cells were treated with 5 μg/mL of Lp(a) or LDL for 6 hours at 37°C. Western blots of cell lysates were performed with an anti-Gpx1 antibody (a), an anti-Prdx6 antibody (b), an anti-Sod1 antibody (c), & an anti-Park7 antibody (d) using an anti-actin antibody as a loading control. Representative blots are shown. Protein levels were normalized against actin & expressed relative to that of untreated cells. Results are expressed as mean ± SEM for pooled triplicate incubations run in quadruplicate. ∗P<0.05, relative to untreated HepG2 cells. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/30596094), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Actin Antibody (mAbGEa) [NB100-74340] -

Western Blot: Actin Antibody (mAbGEa) [NB100-74340] - DenA deneddylase activity.(A) Recombinant human DEN1 & fungal DenA deneddylate a human CUL1-Nedd8 substrate in vitro. SDS-PAGE & subsequent western analysis show cleavage of the substrate (∼60 kDa) producing the C-terminal CUL1 (∼50 kDa) as outlined in experimental procedures. (B) Deneddylation test in a heterologous yeast system. A. nidulans DenA can remove Rub1 from CulD in heterologous expression experiments in S. cerevisiae. DenA expressed as native protein or C-terminally fused w/ a V5/His6 epitope tag. Both variants driven by the inducible GAL1 promoter. CulD, N-terminal-fused w/ the LexA activation domain, expressed under control of the constitutive ADH promoter. A. nidulans proteins expressed in S. cerevisiae wild type & deltacsn5 background. Western analysis w/ antibodies against Rub1 ( alpha-Rub1), the LexA epitope ( alpha-LexA) & the V5 epitope ( alpha-V5) performed. Detection w/ alpha-Rub1 generated two additional signals upon culD expression, representing LexA-CulD & a second CulD pool where LexA unspecifically cleaved off. Both signals disappeared upon co-expression of DenA indicating deneddylation activity (red arrows). The slower migrating signal of alpha-LexA western experiments corresponded to Rub1 modified LexA-CulD. This signal absent when DenA co-expressed. Detection of the V5 tag applied to monitor DenA expression. The neddylated yeast cullin migrating at around 100 kDa not affected by DenA activity. (C) Deneddylation of fungal CulA by CSN & DenA. Whole cell lysates of A. nidulans wild type, deltacsnE & deltadenA probed w/ alpha-CulA. The ratio of neddylated CulA (CulA*N8; ∼106 kDa*) to non-neddylated CulA (∼96 kDa**) calculated from three independent experiments. Membranes reprobed w/ alpha-Actin ( alpha-ACT) for normalization. Image collected & cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pgen.1003275), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Actin Antibody (mAbGEa) [NB100-74340] -

Western Blot: Actin Antibody (mAbGEa) [NB100-74340] - Fibrillar adhesions & FN fibrils are dynamic structures governed by Rab21 & PPFIA1.(a) Living siCTL & siRAB21 ECs incubated w/ IST9 mAb for 30 min, fixed, acid washed & stained. RAB21 silencing impairs ED-A FN endocytosis as revealed by decrease of IST9 punctae co-localizing w/ endosome marker EEA1. Right panels are magnifications of boxed areas in left panels. Data are mean value±s.e.m., n=70 cells per condition pooled from two independent experiments. ***P<0.001. (b) Left panel, WB analysis of PPFIA1, RAB21 & actin on total lysates of siCTL, siPPFIA1 & siPPFIA1+siRAB21 ECs. Right panel, WB analysis of insoluble matrix fraction of ECs extracted w/ DOC buffer. PPFIA1 silencing dramatically reduces amount of DOC-insoluble fraction of endogenous ED-A FN. Of note, simultaneous silencing of Rab21 GTPase (siPPFIA1+siRAB21), which drives integrin endocytosis, rescues defective incorporation of endogenous ED-A FN in DOC-insoluble fraction of siPPFIA1 ECs. (c) Confocal microscopy analysis of patterning of endogenous cellular ED-A FN (green) in fixed confluent ECs. Before fixation, living ECs incubated for 20 min w/ exogenous SNAKA51 (red). ED-A FN polymerizes into a fibrillar network in siCTL, but not in siPPFIA1 ECs. Simultaneous silencing of Rab21 GTPase (siPPFIA1+siRAB21) fully restores ED-A FN polymerization in siPPFIA1 ECs. SNAKA51+ active alpha5 beta1 integrin localizes in fibrillar adhesion in siCTL, but not in siPPFIA1 ECs. Notably, simultaneous Rab21 (siPPFIA1+siRAB21) silencing promotes localization of SNAKA51 in fibrillar adhesion of siPPFIA1 ECs. Data are mean±s.e.m., n=20 cells per condition pooled from two independent experiments. Scale bar, 20 μm (a,c, right), 50μm (c, left). ***P<0.001; Student's t-test. Image collected & cropped by CiteAb from following publication (https://www.nature.com/articles/ncomms13546), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Actin Antibody (mAbGEa) [NB100-74340] -

Western Blot: Actin Antibody (mAbGEa) [NB100-74340] - Effect of the four most common herbal formulas & single herbs on the phosphorylation of myosin light chain (MLC) protein.Briefly, A10 cells were treated with herbal formulas (A) or single herbs (B). Y27632 (Y10; 10 μM) & calyculin A (A50; 50 μg/ml) were used as negative & positive controls. Western blot analysis & staining with anti-phospho-MLC, anti-total-MLC, & anti-beta actin antibodies was then performed. Phospho-MLC, total-MLC, & beta actin were all obtained with their appropriate protein size bands. The relative Phospho-MLC intensity (%) was expressed as [(Phospho-MLC/total-MLC)drug treated/ (Phospho-MLC/total-MLC)cell only x 100%]. The Mean±SEM values for at least three independent experiments along with the representative western blot were performed. Image collected & cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0145109), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Actin Antibody (mAbGEa) [NB100-74340] -

Western Blot: Actin Antibody (mAbGEa) [NB100-74340] - CSN targets DEN1/DenA for degradation in fungal & human cells.(A) Quantitative analysis of repeated western blot experiments displayed the differences in DenA abundance in fungal wild type & deltacsnG cells. DenA levels of the different asexual developmental time points are shown relative to the vegetative (veg.) control for each strain. Anti-Actin was applied as loading control (statistics: 2-way ANOVA; n = 3; *p<0,05, **p<0,01). (B) Xpress-DEN1 was overexpressed in siGFP & siCSN1 human cells & steady state Xpress-DEN1 levels were estimated by western analysis with the alpha-Xpress antibody. Xpress-CSN1 was overexpressed in siCSN1 cells & DEN1 & CSN8 were probed with appropriate antibodies. (C) Xpress-DEN1 was overexpressed in siGFP human cells & the proteasome inhibitor MG132 was added 6 h before cell lysis at a final concentration of 10 µM. Cyclohexamide (CHX) was added in a final concentration of 10 µg/ml (D) Xpress-DEN1 was co-expressed in HeLa cells together with Xpress-CSN1wt, Xpress-CSN1(1–221) or Xpress-CSN1(222–527) in the absence or in the presence of MG132 (right panel), which was added 6 h before cell lysis. Cells were lyzed 24 h after co-transfection & lysates were analyzed by western blot using the alpha-DEN1 antibody (0 = only Xpress-DEN1). Image collected & cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pgen.1003275), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Actin Antibody (mAbGEa) [NB100-74340] -

Western Blot: Actin Antibody (mAbGEa) [NB100-74340] - Analysis of Ia production by engineered S. cerevisiae.(A) Analysis of the synthesis of Ia in S. cerevisiae strains, producing actin-R177K, E270D & E270Q. Yeast strains, producing actin-R177K, E270D or E270Q & transformed with the Ia-containing plasmid (Ia) or the vector alone (Vector), were cultivated in SGal for 20 h at 30°C. Cells were broken by glass beads treatment & analyzed by 32P-ADP-ribosylation in the presence of additionally added purified wild type yeast actin (1 μg). Labeled bands represent modified yeast actin & confirm intracellular production of functionally active Ia by the S. cerevisiae strains. (B) Production of Ia by the wild type S. cerevisiae strain. Wild type yeast strains harboring the Ia-containing plasmid (Ia) or the control vector (vector) were cultivated in glucose-containing liquid medium until OD595 = 0.5. Afterwards, glucose was replaced by galactose & cultivation continued for 9 h at 30°C. Cells were lysed & the resulting extract preparations were ADP-ribosylated in the presence of Ia (+ Ia), TccC3 toxin of P. luminescens [42] (+ TccC3), purified muscle actin (+ alpha-actin) or tested in Western blotting with the anti-actin serum to show equal actin concentrations in the samples. (C, D) Mass spectrometry of actin variants. MALDI-TOF MS of wild type (C) & actin-R177K (D) protein variants isolated from S. cerevisiae. Spectra demonstrate disappearance of R177- & appearance of K177-containing peptide in mass analysis (substituted amino acid residue within identified peptides is shown in red). Image collected & cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0145708), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Actin Antibody (mAbGEa) [NB100-74340] -

Western Blot: Actin Antibody (mAbGEa) [NB100-74340] - alpha5 beta1 regulates ED-A FN secretion & polymerization.(a) Western blot analysis of lysates of ECs control (siCTL) & alpha5 integrin subunit (siITGA5) silenced ECs. Cells were lysed 24 hours after the second siRNA oligofection & proteins were separated by SDS–PAGE & probed for alpha5 integrin subunit or actin (for control purposes). (b) Confocal microscopy analysis of IST9 mAb-labelled endogenous ED-A FN (green) in confluent ECs. ED-A FN polymerizes into a fibrillar network in siCTL, but not in siITGA5 ECs in which it accumulates in the TGN46+ (red) TGN cisternae. The relative amount of fibrillar ED-A FN area was calculated in siCTL & siITGA5 ECs. Data are mean±s.e.m., n=20 cells per condition pooled from two independent experiments. ***P<0.001; Student's t-test. (c) Western blot analysis of soluble ED-A FN released by confluent ECs seeded on Transwell inserts. An equal percentage of apical & basolateral volumes of medium were collected after 72 h of culture, from different wells of siCTL or siITGA5 ECs. Equal amounts of exogenous rabbit IgG were added to samples (spike normalization) for loading control purposes. Quantification of the ratio between apical or basolateral amount of ED-A FN released by siCTL over siITGA5 ECs. alpha5 integrin subunit silencing impairs basolateral, but not apical ED-A FN secretion. Data are mean±s.e.m., n=8 wells per condition pooled from four independent experiments. **P<0.01; Student's t-test. Scale bar, 50 μm (b). Image collected & cropped by CiteAb from the following publication (https://www.nature.com/articles/ncomms13546), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: Actin Antibody (mAbGEa) [NB100-74340] -

Western Blot: Actin Antibody (mAbGEa) [NB100-74340] - Effect of the four most common herbal formulas & single herbs on the phosphorylation of myosin light chain (MLC) protein.Briefly, A10 cells were treated with herbal formulas (A) or single herbs (B). Y27632 (Y10; 10 μM) & calyculin A (A50; 50 μg/ml) were used as negative & positive controls. Western blot analysis & staining with anti-phospho-MLC, anti-total-MLC, & anti-beta actin antibodies was then performed. Phospho-MLC, total-MLC, & beta actin were all obtained with their appropriate protein size bands. The relative Phospho-MLC intensity (%) was expressed as [(Phospho-MLC/total-MLC)drug treated/ (Phospho-MLC/total-MLC)cell only x 100%]. The Mean±SEM values for at least three independent experiments along with the representative western blot were performed. Image collected & cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0145109), licensed under a CC-BY license. Not internally tested by Novus Biologicals.