Anti-GRP78/BiP (ET-21) antibody produced in rabbit
SIGMA/G9043 - IgG fraction of antiserum, buffered aqueous solution
Synonym: Anti-78-kDa Glucose-Regulating Protein; Anti-Immunoglobulin Heavy Chain Binding Protein
MDL Number: MFCD01865530
Product Type: Chemical
Immunoblotting
HeLa cell extracts (Lane A, B) and recombinant hamster GRP78 protein (Lane C) were separated on SDS-PAGE, blotted with Anti-GRP78/BiP (ET-21) (Cat. No. G9043) at a 1:3,000 dilution and developed using Anti-Rabbit IgG, Peroxidase (Cat. No. A0545) and a chemiluminescent substrate. An additional band at 73 kDa may represent a GRP78 degradation product. Lane B: Inhibition of staining with GRP78 immunizing peptide (20 μg/mL).
HeLa cell extracts (Lane A, B) and recombinant hamster GRP78 protein (Lane C) were separated on SDS-PAGE, blotted with Anti-GRP78/BiP (ET-21) (Cat. No. G9043) at a 1:3,000 dilution and developed using Anti-Rabbit IgG, Peroxidase (Cat. No. A0545) and a chemiluminescent substrate. An additional band at 73 kDa may represent a GRP78 degradation product. Lane B: Inhibition of staining with GRP78 immunizing peptide (20 μg/mL).
Enhanced Validation-By Independent Antibodies Immunofluorescence. 3T3 cells were fixed and permeabilized with methanol followed by acetone. Fixed cells were stained with 1:1,000 Anti-GRP78/BiP (GL-19) antibody produced in Rabbit (Cat. No. G8918) (A). The antibody developed using Goat Anti-Rabbit IgG, FITC conjugate antibody (Cat. No. F9887), and 1:1,000 Anti-GRP78/BiP (ET-21) antibody produced in Rabbit (Cat. No. G9043) (B). The antibody was developed using Goat Anti-Rabbit IgG, FITC conjugate antibody (Cat. No. F9887). Results Two Anti-GRP78/BiP antibodies, G8918 (A) and G9043 (B), target different regions of GRP78/BiP show similar staining profiles between the two antibodies, demonstrating Independent Antibody Verification.
Immunohistochemistry (Paraffin) Formalin Fixed Paraffin Embedded (FFPE) human thyroid (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:1,000 dilution of Cat. No. G9043, Anti-GRP78/BiP (ET-21) antibody. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous staining was observed in follicular epithelium of human thyroid tissue.
Immunohistochemistry (Paraffin) Formalin Fixed Paraffin Embedded (FFPE) human cerebral cortex (A) and Negative control (no primary) (B) tissue sections were prepared using heat-induced epitope retrieval (HIER). Immunostaining was performed using a 1:1,000 dilution of Cat. No. G9043, Anti-GRP78/BiP (ET-21) antibody. Reactivity was detected using a Goat Anti-Rabbit IgG and HRP-DAB. Cytoplasmic/membranous/nuclear staining was observed in neurons and glial cells of human cerebral cortex tissue sections.
Immunocytochemistry Methanol/acetone fixed cultured mouse fibroblast NIH3T3 cell line stained with 1:1,000 Anti-GRP78/BiP (ET-21) antibody produced in Rabbit (Cat. No. G9043) followed by 1:75 Anti-Rabbit IgG (whole molecule)-FITC antibody produced in Goat (Cat. No. F9887).
| antibody form | IgG fraction of antiserum |
| antibody product type | primary antibodies |
| biological source | rabbit |
| clone | polyclonal |
| conjugate | unconjugated |
| enhanced validation | independent Learn more about Antibody Enhanced Validation  |
| form | buffered aqueous solution |
| mol wt | antigen 78 kDa |
| Quality Level | 200 |
| shipped in | dry ice |
| species reactivity | mouse, chicken, Xenopus, hamster, human, rat |
| storage temp. | −20°C |
| target post-translational modification | unmodified |
| technique(s) | immunocytochemistry: 1:1,000 using methanol/acetone fixed cultured mouse fibroblast NIH3T3 cell line |
| immunohistochemistry: 1:1,000 using human cerebral cortex and human thyroid gland tissue sections | |
| western blot: 1:3,000 using whole cell extract of human epitheloid carcinoma HeLa cell line | |
| UniProt accession no. | P11021  |
| Application: | Anti-GRP78/BiP (ET-21) antibody produced in rabbit is suitable for use as a primary antibody: • for immunofluorescence staining of frozen heart tissue to examine whether ER stress signaling activation occurs in cardiomyocytes • for immunohistochemistry on Paraffin embedded sections of human cerebral cortex and human thyroid gland tissue sections. • to detect Bip by immunoblotting of protein extract from WT AB strain of zebrafish (Danio rerio) • at a working dilution of 1:1000 to detect GRP78 in extract of prostate cancer cells after androgen deprivation therapy It is also suitable for immunoblotting at a working dilution of 1:3000 using human epitheloid carcinoma HeLa whole cell extract and for immunocytochemistry at a working dilution of 1:1000 using mouse fibroblasts NIH3T3 cells. |
| Biochem/physiol Actions: | GRP78/BiP levels are elevated in Alzheimer′s disease brains and the decreased expression of GRP78/BiP is found associated with missense mutations in presenilin-1 (PS-1). |
| Biochem/physiol Actions: | The GRP78 (78-kDa glucose-regulated protein) is a member of the heat shock proteins Hsp70 required for cell viability. It is a Ca2+-binding molecular chaperone that is localized to the endoplasmic reticulum (ER). It plays a key role in proper glycosylation, folding and assembly of newly synthesized membrane bound or secretory proteins. It also facilitates retention of mutant or defective proteins that are improperly folded and prevents translocation of such proteins from the cytosol to the ER lumen. The protein is induced under conditions of stress such as oxidative stress, chemical toxicity, glycosylation and hypoxia. It plays a crucial role in the maintenance of cell homeostasis and the prevention of apoptosis and acts as a biomarker of hypoglycemia. It has a neuroprotective function in neurons exposed to glutamate exotoxicity and oxidative stress. |
| Disclaimer: | Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals. |
| Packaging: | 200 μL in glass insert |
| Physical form: | Solution in 0.01 M phosphate buffered saline, pH 7.4, containing 15 mM sodium azide. |
| Hazard Codes | Xn |
| Risk Statements | 21/22 |
| Safety Statements | 36/37-60 |
| RIDADR | NONH for all modes of transport |
| WGK Germany | nwg |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | −20°C |
| UNSPSC | 12352203 |