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

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

Detection of Human GRP78/HSPA5 antibody by Western Blot.

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 antibody by Simple WesternTM.

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

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

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

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

Application
Recommended Usage

Simple Western

10 µg/mL
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

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

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: GRP78/HSPA5

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.

Long Name

75 KD Glucose Regulated Protein

Alternate Names

BIP, HSP70-5, HSPA5

Entrez Gene IDs

3309 (Human); 14828 (Mouse); 25617 (Rat)

Gene Symbol

HSPA5

UniProt

Additional GRP78/HSPA5 Products

Product Documents for Human GRP78/HSPA5 Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human GRP78/HSPA5 Antibody

For research use only

Citations for Human GRP78/HSPA5 Antibody

Customer Reviews for Human GRP78/HSPA5 Antibody (1)

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  • Human GRP78/HSPA5 Antibody
    Name: Byron Baron
    Application: Western Blot
    Sample Tested: MCF7 (breast)
    Species: Human
    Verified Customer | Posted 11/03/2016
    Marker, 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
    Human GRP78/HSPA5 Antibody MAB4846

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