GRP78 (Glucose-regulated protein 78 kDa; also BiP and HSPA5) is a 72 kDa member of the heat shock protein 70 family of proteins. Intracellularly, GRP78 is an endoplasmic reticulum chaperone that participates in protein folding; extracellularly, it induces IL-10 production from T cells and interacts with Cripto to block TGF-beta signaling. Human GRP78 precursor is 654 amino acids (aa) in length. It contains an 18 aa signal sequence and a 636 aa mature region that shows a hydantoinase A region (aa 145‑245) and a C-terminal KDEL motif that is present on intracellular GRP78, but absent on secreted GRP78. There is alternative splicing in the signal sequence (aa 1‑10), and multiple single aa substitituion. Over aa 1‑654, human GRP78 is more than 97% aa identical to mouse and rat GRP78.
Human GRP78/HSPA5 Antibody (474421)
R&D Systems | Catalog # MAB4846
Scientific images may be resized, cropped, or adjusted for brightness or contrast for presentation purposes. These updates are limited to presentation and do not affect the underlying scientific interpretation of the data. Where available, additional source data may be provided upon request.
Key Product Details
Species Reactivity
Validated:
Human
Cited:
Human, Fish - Danio rerio (Zebrafish)
Applications
Validated:
Western Blot, Simple Western
Cited:
Western Blot, Immunocytochemistry, Immunoprecipitation
Label
Unconjugated
Antibody Source
Monoclonal Mouse IgG2B Clone # 474421
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Product Specifications
Immunogen
E. coli-derived recombinant human GRP78
Met1-Leu654
Accession # P11021
Met1-Leu654
Accession # P11021
Specificity
Detects endogenous human GRP78 at 78 kDa in Western blots.
Clonality
Monoclonal
Host
Mouse
Isotype
IgG2B
Scientific Data Images for Human GRP78/HSPA5 Antibody (474421)
Detection of Human GRP78/HSPA5 by Western Blot.
Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line, Jurkat human acute T cell leukemia cell line, and MCF-7 human breast cancer cell line. PVDF membrane was probed with 1 µg/mL of Human GRP78/HSPA5 Monoclonal Antibody (Catalog # MAB4846) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for GRP78/HSPA5 at approximately 78 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 2.
Detection of Human GRP78/HSPA5 by Simple WesternTM.
Simple Western lane view shows lysates of Jurkat human acute T cell leukemia cell line, loaded at 0.2 mg/mL. A specific band was detected for GRP78/HSPA5 at approximately 72 kDa (as indicated) using 10 µg/mL of Mouse Anti-Human GRP78/HSPA5 Monoclonal Antibody (Catalog # MAB4846). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.*Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody.
Detection of GRP78/HSPA5 by Western Blot
Western blot validation of identified proteins to confirm the expression trend as inferred from 2DE. Protein levels were normalized against GAPDH. Error bars represent S.E.M. * represents (p < 0.05) and ** represents (p < 0.01). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/22357162), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of GRP78/HSPA5 by Western Blot
Extracellular vesicle secretion induced by Sorafenib and their miRNA content. (A) Particle size and concentration analysis of Large, Small, and Very Small EVs obtained at 6 and 24 h. (B) Expression of EV markers and cellular contaminants in Large, Small, and Very Small EVs and cell lysates obtained at 24 h after Sorafenib treatment. Total lane protein content was used as the loading control. (C) Representative images of cryo-EM of Small and Very Small EVs obtained at 6 and 24 h from the control and Sorafenib-treated cells. (D) Assessment of EV size (nm) in the cryo-EM images. (E) Number of EVs quantified in the cryo-EM images. (F) miRNA expression in the three fractions of EVs at 6 h. (G) miRNA expression in the three fractions of EVs at 24 h. Fold-change values were calculated between Sorafenib and the control treated samples. Results are expressed as the mean ± SEM of six independent experiments. Ns, non-significant; * p ≤0.05, ** p ≤ 0.01, *** p ≤ 0.001, and **** p ≤ 0.0001 between the miRNA expression in the control and Sorafenib derived EVs. Multiple comparison test statistics are expressed with lower case a–i. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36078082), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of GRP78/HSPA5 by Western Blot
Extracellular vesicle secretion induced by Sorafenib and their miRNA content. (A) Particle size and concentration analysis of Large, Small, and Very Small EVs obtained at 6 and 24 h. (B) Expression of EV markers and cellular contaminants in Large, Small, and Very Small EVs and cell lysates obtained at 24 h after Sorafenib treatment. Total lane protein content was used as the loading control. (C) Representative images of cryo-EM of Small and Very Small EVs obtained at 6 and 24 h from the control and Sorafenib-treated cells. (D) Assessment of EV size (nm) in the cryo-EM images. (E) Number of EVs quantified in the cryo-EM images. (F) miRNA expression in the three fractions of EVs at 6 h. (G) miRNA expression in the three fractions of EVs at 24 h. Fold-change values were calculated between Sorafenib and the control treated samples. Results are expressed as the mean ± SEM of six independent experiments. Ns, non-significant; * p ≤0.05, ** p ≤ 0.01, *** p ≤ 0.001, and **** p ≤ 0.0001 between the miRNA expression in the control and Sorafenib derived EVs. Multiple comparison test statistics are expressed with lower case a–i. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36078082), licensed under a CC-BY license. Not internally tested by R&D Systems.Applications for Human GRP78/HSPA5 Antibody (474421)
Application
Recommended Usage
Simple Western
10 µg/mL
Sample: Jurkat human acute T cell leukemia cell line
Sample: Jurkat human acute T cell leukemia cell line
Western Blot
1 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line, Jurkat human acute T cell leukemia cell line, and MCF-7 human breast cancer cell line
Sample: HeLa human cervical epithelial carcinoma cell line, Jurkat human acute T cell leukemia cell line, and MCF-7 human breast cancer cell line
Reviewed Applications
Read 1 review rated 4 using MAB4846 in the following applications:
Formulation, Preparation, and Storage
Purification
Protein A or G purified from hybridoma culture supernatant
Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.
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Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
- 12 months from date of receipt, -20 to -70 °C as supplied.
- 1 month, 2 to 8 °C under sterile conditions after reconstitution.
- 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Calculators
Background: GRP78/HSPA5
Long Name
75 KD Glucose Regulated Protein
Alternate Names
BIP, HSP70-5, HSPA5
Gene Symbol
HSPA5
UniProt
Additional GRP78/HSPA5 Products
Product Documents for Human GRP78/HSPA5 Antibody (474421)
Certificate of Analysis
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Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Human GRP78/HSPA5 Antibody (474421)
For research use only
Related Research Areas
Citations for Human GRP78/HSPA5 Antibody (474421)
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Application: Western BlotSample Tested: MCF7 (breast)Species: HumanVerified Customer | Posted 11/03/2016Marker, MCF7 (breast), 231 (breast), Colo320 (colon), HeLa (cervix) (25ug protein per well)Primary mouse anti GRP78 antibody diluted 1:1000 in TBS. Membrane incubation for 2 hours at RT Secondary IRDye 800CW goat anti mouse antibody diluted 1:10000 in TBS. Membrane incubation for 1 hour at RT Detection by Li-Cor Odyssey
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- Cellular Response to Hypoxia Protocols
- R&D Systems Quality Control Western Blot Protocol
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
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