GRP78/HSPA5 Antibody - BSA Free

Immunocytochemistry/ Immunofluorescence: GRP78/HSPA5 Antibody - BSA Free [NBP1-06274]

Immunocytochemistry/Immunofluorescence: GRP78/HSPA5 Antibody [NBP1-06274] - NIH-3T3 cells were fixed and permeabilized for 10 minutes using -20C MeOH. The cells were incubated with anti-GRP78/HSPA5 at 2 ug/mL overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Western Blot: GRP78/HSPA5 AntibodyBSA Free [NBP1-06274]

Western Blot: GRP78/HSPA5 Antibody [NBP1-06274] - Total protein from human HeLa and A431 cells, mouse 3T3 cells and rat PC12 cells was separated on a 7.5% gel by SDS-PAGE, transferred to PVDF membrane and blocked in 5% non-fat milk in TBST. The membrane was probed with 1.0 ug/mL anti-GPR78 in blocking buffer and detected with an anti-rabbit HRP secondary antibody using chemiluminescence.

Immunocytochemistry/ Immunofluorescence: GRP78/HSPA5 Antibody - BSA Free [NBP1-06274]

Immunocytochemistry/Immunofluorescence: GRP78/HSPA5 Antibody [NBP1-06274] - 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-GRP78/HSPA5 at 5 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Alpha tubulin (DM1A) NB100-690 was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Flow Cytometry: GRP78/HSPA5 Antibody - BSA Free [NBP1-06274]

Flow Cytometry: GRP78/HSPA5 Antibody [NBP1-06274] - An intracellular stain was performed on NIH3T3 cells with GRP78/HSPA5 Antibody NBP1-06274AF488 (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 5 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 488.

Western Blot: GRP78/HSPA5 AntibodyBSA Free [NBP1-06274]

Western Blot: GRP78/HSPA5 Antibody [NBP1-06274] - Detection of Bip/Grp78 on HeLa whole cell extracts using NBP1-06274.

Flow Cytometry: GRP78/HSPA5 Antibody - BSA Free [NBP1-06274]

Flow Cytometry: GRP78/HSPA5 Antibody [NBP1-06274] - Analysis using Alexa Fluor (R) 488 conjugate of NBP1-06274. An intracellular stain was performed on HeLa cells with GPR78/HSPA5 antibody NBP1-06274 (blue) and a matched isotype control NBP2-24893 (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. 1 ug of antibody was added to 100 uL of staining buffer and cells were incubated for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 488.

Flow Cytometry: GRP78/HSPA5 Antibody - BSA Free [NBP1-06274]

Flow Cytometry: GRP78/HSPA5 Antibody [NBP1-06274] - An intracellular stain was performed on Jurkat cells with Alexa Fluor 647 conjugate of GPR78/HSPA5 antibody NBP1-06274AF647 (blue) and a matched isotype control NBP2-24893AF647 (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 2 ug/mL for 30 minutes at room temperature. Both antibodies were conjugated to Alexa Fluor 647.

Flow Cytometry: GRP78/HSPA5 Antibody - BSA Free [NBP1-06274]

Flow Cytometry: GRP78/HSPA5 Antibody [NBP1-06274] - An intracellular stain was performed on HeLa with NBP1-06274 and a matched isotype control. 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 Rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody, Dylight 550.

Western Blot: GRP78/HSPA5 Antibody - BSA Free [NBP1-06274] -

Western Blot: GRP78/HSPA5 Antibody - BSA Free [NBP1-06274] - Hotair promotes the expression of GRP78 & PD-L1, mediates the proliferation, & helps to regulate the T cell-mediated immune responses in Tu212 & Hep-2 cells. Western blot analysis of GRP78 & PD-L1 after mi-NC or has-miR-30a-5p & vehicle or Hotair transfection in Hep-2 cells (a). The relative expression level of GRP78 & PD-L1 in Hep-2 cells (b). (c–e) The cell viability, apoptosis, & colony formation after mi-NC or has-miR-30a-5p & vehicle or Hotair transfection in Hep-2 cells. (f–h) T cell killing assay of target Tu212 & Hep-2 cells cocultured with PBMC, & ELISA detects the concentration of cytokines (g) IFN-gamma & (h) IL-2. The targeted Tu212 & Hep-2 cells after 24 h transfected with mi-NC or has-miR-30a-5p & vehicle or Hotair were cocultured with PBMC, & the cytotoxicity was measured by the LDH release assay. Data are mean ± SD of at least three independent experiments; ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/35419461), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: GRP78/HSPA5 Antibody - BSA Free [NBP1-06274] -

Western Blot: GRP78/HSPA5 Antibody - BSA Free [NBP1-06274] - Analysis of protein levels involved in ER stress & Ca2+ homeostasis & processing of mRNA XBP1 in control & patients-derived fibroblasts.(A) Equal amounts from controls & patients were loaded (50 μg of total cell lysates) & subjected to Western Blot with anti-Herp, anti-Grp78 & anti-IP3R1 antibodies. We used anti-Hsp60 antibody to ensure equal amounts of protein loaded in each lane. This result is representative of three independent experiments. Protein quantification was performed by laser densitometry. The ratios between proteins/Hsp60 for each cell line were calculated to determine the expression fold-change relative to control. (B) Data represent mean ± standard deviation of three independent experiments. (C) Equal amounts from controls were loaded (50 μg of total cell lysates) & subjected to Western Blot with anti-Herp & anti-Grp78 antibodies. We used anti-GAPDH antibody to ensure equal amounts of protein loaded in each lane. This result is representative of two independent experiments. (D) Equal amounts from control & patients were loaded (50 μg of total cell lysates) & subjected to Western Blot with anti-phospho-PERK antibody. We used anti-GAPDH antibody to ensure equal amounts of protein loaded in each lane. This result is representative of two independent experiments. (E) RT-PCR analysis of the processing of mRNA XBP1 transcription factor. Tm: tunicamycin; u: XBP1 unspliced form; s: XBP1 spliced form. Image collected & cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0150357), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Western Blot: GRP78/HSPA5 Antibody - BSA Free [NBP1-06274] -

Western Blot: GRP78/HSPA5 Antibody - BSA Free [NBP1-06274] - UPR Components Activated in A beta-Expressing Flies Are Induced Even Further by Glut1 Overexpression(A) Western blot of eIF2 phosphorylation levels in heads of A beta- & A betaGlut1-expressing flies (+RU) & in controls (-RU), showing no significant difference. Bottom: plotted as means ± SEM (n = 3). Top: a representative gel from the same samples.(B) Grp78 mRNA levels in heads of 18-day-old flies expressing A beta or A beta Glut1 in neurons (+RU) & uninduced controls (-RU), measured by qPCR (relative to eIF1A), plotted as means ± SEM. Genotypes: UAS A beta; elavGS, UAS A beta/UAS Glut1; elavGS.(C) Quantification of GFP fluorescence in fly brains expressing an Xbp1GFP splicing reporter, plotted as means ± SEM (n = 6–13). Genotypes: elavGS/UAS-Xbp1GFP, UAS A beta; elavGS/UAS-Xbp1GFP, UAS A beta/UAS Glut1; elavGS/UAS-Xbp1GFP.(D) Spliced Xbp1 mRNA levels in heads of 18-day-old flies expressing A beta or A beta Glut1 in neurons (+RU) & uninduced controls (-RU), measured by qPCR (relative to eIF1A), plotted as means ± SEM(E) Western blot of Grp78 in 14-day-old flies of the same genotypes, plotted below as means ± SEM (n = 6–16). The image is a representative gel of the same samples. Genotypes: UAS A beta; elavGS, UAS A beta/UAS Glut1; elavGS.∗p ≤ 0.05; ∗∗p ≤ 0.01, by ANOVA. See also Figure S2. Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/27524482), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

Immunocytochemistry/ Immunofluorescence: GRP78/HSPA5 Antibody - BSA Free [NBP1-06274] -

Immunocytochemistry/ Immunofluorescence: GRP78/HSPA5 Antibody - BSA Free [NBP1-06274] - C. elegans granulins impair organismal fitness & resistance to ER stress.(A) Wild-type (N2) & pgrn-1(-) animals with & without granulin expression were subjected to ER stress with tunicamycin (5 μg / ml). The fraction developing to L4 stage was quantified (n = 50, 3 biological replicates). (B) Wild-type (N2) & pgrn-1(-) animals with & without C. elegans progranulin over-expression (OE) were subjected to ER stress with tunicamycin (5 μg / ml). The fraction developing to L4 stage was quantified (n = 50, 3 biological replicates). (C) Total worm lysates from synchronized day 1 adult granulin-expressing animals were immunoblotted with an anti-HSP-4/BiP antibody (3 biological replicates). Anti-actin was used as a loading control. (D) Wild-type & pgrn-1(-) animals with & without granulin expression were staged as embryos. Animals were scored for development to L4 stage (n = 50, 12 biological replicates). (E) Measurement of body length at day 1 adulthood (n = 12). (F) Measurement of short-term associative learning (three biological replicates). The glutamate receptor mutant nmr-1(ak4) was used as a positive control. Throughout, error bars show mean ± SEM, one or two-way ANOVA with post-hoc Tukey multiple comparisons test. Comparisons are to wild-type unless otherwise indicated (*P<0.05, ***P<0.001, ****P<0.0001, ns = not significant, wt = wild-type). Image collected & cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31398187), licensed under a CC-BY license. Not internally tested by Novus Biologicals.