Anti-GRP78 BiP antibody ab21685 is a rabbit polyclonal antibody that is used in GRP78 BiP western blotting, IHC and immunofluorescence. Suitable for human, mouse and rat samples.
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- Iranian journal of basic medical sciences 27:114-1212024Chronic stress-induced anxiety-like behavior, hippocampal oxidative, and endoplasmic reticulum stress are reversed by young plasma transfusion in aged adult rats.Applications:Unspecified applicationReactive species:Unspecified reactive speciesArshad Ghaffari-Nasab,Gonja Javani,Fereshteh Farajdokht,Mohammad Reza Alipour,Gisou MohaddesPubMed 38164475
- The Journal of clinical investigation 134:2024Hypomorphic variants of SEL1L-HRD1 ER-associated degradation are associated with neurodevelopmental disorders.Applications:Unspecified applicationReactive species:Unspecified reactive speciesHuilun H Wang,Liangguang L Lin,Zexin J Li,Xiaoqiong Wei,Omar Askander,Gerarda Cappuccio,Mais O Hashem,Laurence Hubert,Arnold Munnich,Mashael Alqahtani,Qi Pang,Margit Burmeister,You Lu,Karine Poirier,Claude Besmond,Shengyi Sun,Nicola Brunetti-Pierri,Fowzan S Alkuraya,Ling QiPubMed 37943610
- The Journal of clinical investigation 134:2024Biallelic Cys141Tyr variant of SEL1L is associated with neurodevelopmental disorders, agammaglobulinemia, and premature death.Applications:Unspecified applicationReactive species:Unspecified reactive speciesDenisa Weis,Liangguang L Lin,Huilun H Wang,Zexin Jason Li,Katarina Kusikova,Peter Ciznar,Hermann M Wolf,Alexander Leiss-Piller,Zhihong Wang,Xiaoqiong Wei,Serge Weis,Katarina Skalicka,Gabriela Hrckova,Lubos Danisovic,Andrea Soltysova,Tingxuan T Yang,René Günther Feichtinger,Johannes A Mayr,Ling QiPubMed 37943617
- Molecular and cellular biochemistry :2023HRD1 reduction promotes cholesterol-induced vascular smooth muscle cell phenotypic change via endoplasmic reticulum stress.Applications:Unspecified applicationReactive species:Unspecified reactive speciesLinli Wang,Zhitao Ren,Lin Wu,Ximei Zhang,Min Wang,Haiming Niu,Xuemin He,Heting Wang,Yanming Chen,Guojun Shi,Xiaoxian QianPubMed 38145449
- Molecular medicine (Cambridge, Mass.) 29:1632023Sustained activation of 12/15 lipoxygenase (12/15 LOX) contributes to impaired renal recovery post ischemic injury in male SHR compared to females.Applications:Unspecified applicationReactive species:Unspecified reactive speciesRiyaz Mohamed,Jennifer C SullivanPubMed 38049738
- Science advances 9:eadi73472023Copper drives prion protein phase separation and modulates aggregation.Applications:Unspecified applicationReactive species:Unspecified reactive speciesMariana Juliani do Amaral,Satabdee Mohapatra,Aline Ribeiro Passos,Taiana Sousa Lopes da Silva,Renato Sampaio Carvalho,Marcius da Silva Almeida,Anderson Sá Pinheiro,Susanne Wegmann,Yraima CordeiroPubMed 37922348
- FASEB journal : official publication of the Federation of American Societies for Experimental Biology 37:e232742023GRP78 acts as a cAMP/PKA signaling modulator through the MC4R pathway in porcine embryonic development.Applications:Unspecified applicationReactive species:Unspecified reactive speciesGeun Heo,Song-Hee Lee,Ji-Dam Kim,Gyu-Hyun Lee,Jae-Min Sim,Dongjie Zhou,Jing Guo,Xiang-Shun CuiPubMed 37917004
- Frontiers in cell and developmental biology 11:12385462023Knockdown of Y-box binding protein 1 induces autophagy in early porcine embryos.Applications:Unspecified applicationReactive species:Unspecified reactive speciesWen-Jie Jiang,Song-Hee Lee,Geun Heo,Hak Jae Chung,Eun Seok Cho,Soo Jin Sa,Shinichi Hochi,Xiang-Shun CuiPubMed 37965572
- Experimental and therapeutic medicine 26:5752023IGF2BP2‑dependent STIM1 inhibition protects against LPS‑induced pneumonia by alleviating endoplasmic reticulum stress and the inflammatory response.Applications:Unspecified applicationReactive species:Unspecified reactive speciesWei Zhou,Qigang Dai,Ning Su,Zhihui Liu,Jinxing HuPubMed 38023363
- Molecular neurobiology 61:2313-23352023Sevoflurane Exposure Induces Neuronal Cell Ferroptosis Initiated by Increase of Intracellular Hydrogen Peroxide in the Developing Brain via ER Stress ATF3 Activation.Applications:Unspecified applicationReactive species:Unspecified reactive speciesLiheng Kang,Meihua Piao,Nan Liu,Wanping Gu,Chunsheng FengPubMed 37874483
Key facts
- Isotype
IgG
- Host species
Rabbit
- Storage buffer
pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: 98.98% PBS, 1% BSA- Form
Liquid
- Clonality
Polyclonal
Immunogen
- The exact immunogen used to generate this antibody is proprietary information.
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Reactivity data
Select an application| IHC-P | ICC/IF | WB | |
|---|---|---|---|
| Human | Tested | Tested | Tested |
| Mouse | Expected | Expected | Tested |
| Rat | Expected | Expected | Tested |
| African green monkey | Predicted | Predicted | Predicted |
| Chinese hamster | Expected | Expected | Tested |
| Dog | Predicted | Predicted | Predicted |
| Pig | Predicted | Predicted | Predicted |
IHC-P
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info 1 µg/mL | Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
ExpectedExpected
| Species | Dilution info | Notes |
|---|---|---|
Species Mouse | Dilution info 1 µg/mL | Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
Species Chinese hamster | Dilution info 1 µg/mL | Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
Species Rat | Dilution info 1 µg/mL | Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
PredictedPredicted
| Species | Dilution info | Notes |
|---|---|---|
Species Dog, Pig, African green monkey | Dilution info - | Notes - |
ICC/IF
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info 1-5 µg/mL | Notes We recommend using Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) secondary antibody. |
ExpectedExpected
| Species | Dilution info | Notes |
|---|---|---|
Species Mouse | Dilution info 1-5 µg/mL | Notes We recommend using Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) secondary antibody. |
Species Chinese hamster | Dilution info 1-5 µg/mL | Notes We recommend using Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) secondary antibody. |
Species Rat | Dilution info 1-5 µg/mL | Notes We recommend using Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) secondary antibody. |
PredictedPredicted
| Species | Dilution info | Notes |
|---|---|---|
Species Dog, Pig, African green monkey | Dilution info - | Notes - |
WB
TestedTested
| Species | Dilution info | Notes |
|---|---|---|
Species Human | Dilution info 1 µg/mL | Notes - |
Species Chinese hamster | Dilution info 1 µg/mL | Notes - |
Species Mouse | Dilution info 1 µg/mL | Notes - |
Species Rat | Dilution info 1 µg/mL | Notes - |
PredictedPredicted
| Species | Dilution info | Notes |
|---|---|---|
Species Dog, Pig, African green monkey | Dilution info - | Notes - |
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Target data
Function
Endoplasmic reticulum chaperone that plays a key role in protein folding and quality control in the endoplasmic reticulum lumen (PubMed:2294010, PubMed:23769672, PubMed:23990668, PubMed:28332555). Involved in the correct folding of proteins and degradation of misfolded proteins via its interaction with DNAJC10/ERdj5, probably to facilitate the release of DNAJC10/ERdj5 from its substrate (By similarity). Acts as a key repressor of the ERN1/IRE1-mediated unfolded protein response (UPR) (PubMed:1550958, PubMed:19538957). In the unstressed endoplasmic reticulum, recruited by DNAJB9/ERdj4 to the luminal region of ERN1/IRE1, leading to disrupt the dimerization of ERN1/IRE1, thereby inactivating ERN1/IRE1 (By similarity). Accumulation of misfolded protein in the endoplasmic reticulum causes release of HSPA5/BiP from ERN1/IRE1, allowing homodimerization and subsequent activation of ERN1/IRE1 (By similarity). Plays an auxiliary role in post-translational transport of small presecretory proteins across endoplasmic reticulum (ER). May function as an allosteric modulator for SEC61 channel-forming translocon complex, likely cooperating with SEC62 to enable the productive insertion of these precursors into SEC61 channel. Appears to specifically regulate translocation of precursors having inhibitory residues in their mature region that weaken channel gating. May also play a role in apoptosis and cell proliferation (PubMed:26045166).(Microbial infection) Plays an important role in viral binding to the host cell membrane and entry for several flaviruses such as Dengue virus, Zika virus and Japanese encephalitis virus (PubMed:33432092, PubMed:15098107, PubMed:28053106). Acts as a component of the cellular receptor for Dengue virus serotype 2/DENV-2 on human liver cells (PubMed:15098107).
Alternative names
Endoplasmic reticulum chaperone BiP, 78 kDa glucose-regulated protein, Binding-immunoglobulin protein, Heat shock protein 70 family protein 5, Heat shock protein family A member 5, Immunoglobulin heavy chain-binding protein, GRP-78, BiP, HSP70 family protein 5, HSPA5, GRP78
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Anti-GRP78 BiP antibody ab21685 is a rabbit polyclonal antibody that is used in GRP78 BiP western blotting, IHC and immunofluorescence. Suitable for human, mouse and rat samples.
- Tried and trusted by researchers since 2006
Key facts
- Isotype
IgG
- Host species
Rabbit
- Storage buffer
pH: 7.4
Preservative: 0.02% Sodium azide
Constituents: 98.98% PBS, 1% BSA- Form
Liquid
- Clonality
Polyclonal
- Immunogen
- The exact immunogen used to generate this antibody is proprietary information.
- Purification technique
Affinity purification Immunogen
- Specificity
Replenishment batches of our polyclonal antibody, ab21685 are tested in WB. Previous batches were additionally validated in ICC/IF and IHC-P. These applications are still expected to work and are covered by our Abpromise guarantee. You may also be interested in our alternative recombinant antibody, ab108613.
- Concentration
Storage
- Shipped at conditions
Blue Ice
- Appropriate short-term storage duration
1-2 weeks
- Appropriate short-term storage conditions
+4°C
- Appropriate long-term storage conditions
-20°C
- Aliquoting information
Upon delivery aliquot
- Storage information
Avoid freeze / thaw cycle
Notes
Stimulation may be required to allow detection of the target protein due to low levels of endogenous expression in some samples. Please see images below for recommended treatment conditions and positive controls.
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Supplementary info
This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
GRP78 also known as BiP or HSPA5 is a protein that plays an important role in protein folding and assembly within the endoplasmic reticulum (ER). It weighs approximately 78 kDa hence the name GRP78. This chaperone protein binds to hydrophobic regions of nascent polypeptides to prevent aggregation and misfolding. GRP78 is expressed in high levels in the ER of cells where it monitors cellular stress and aids in maintaining ER homeostasis.
- Biological function summary
GRP78/BiP is important in the unfolded protein response (UPR) a cellular stress response related to the ER. It is part of a complex that manages protein load inside the ER by regulating the fold of nascent proteins and interacting with other ER stress sensors like IRE1 PERK and ATF6. This function is essential for maintaining proper protein conformation in the ER especially during physiological stress ensuring that only correctly folded proteins proceed to the Golgi apparatus.
- Pathways
GRP78/BiP significantly affects the UPR and apoptosis pathways. By controlling protein folding and quality in the ER GRP78 helps prevent cell death under stress conditions. It works closely with other proteins like ATF6 which activates stress response genes. In normal conditions GRP78 keeps stress transducers inactivated but during stress it dissociates allowing UPR signaling to occur.
- Associated diseases and disorders
GRP78/BiP has been implicated in cancer and neurodegenerative diseases. Overexpression of GRP78 is associated with tumor proliferation and poor prognosis in various cancers as cancer cells heavily rely on its protein-folding capacity to survive. Additionally in neurodegenerative diseases misfolded proteins can accumulate leading GRP78 to attempt counteracting these toxic aggregations highlighting its connection with proteins like tau and amyloid-beta in diseases such as Alzheimer's.
Product promise
Tested
We have tested this species and application combination and it works. It is covered by our product promise.
Expected
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
Predicted
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
Not recommended
We do not recommend this combination. It is not covered by our product promise.
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In the unlikely event of one of our products not working as expected, you are covered by our product promise.
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10 product images
Image from Friebe S et al., PLoS One. 2015;10(3):e0119864. Fig 3.; doi: 10.1371/journal.pone.0119864. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/Immunocytochemistry/ Immunofluorescence - Anti-GRP78 BiP antibody (ab21685)
Localization of TEM8 and CMG2 glycosylation mutants.
A) Immunofluorescence of transiently transfected HeLa cells. Cells were transfected for 48h with the respective cDNAs. Cells were fixed, permeabilized and stained for TEM8-HA, endogenous BiP and Hoechst. Scalebars represent 10 μm.
B) Immunofluorescence of transiently transfected HeLa cells. Cells were transfected for 48h with the respective cDNAs. Cells were fixed, permeabilized and stained for CMG2-V5, endogenous BiP and Hoechst. Scalebars represent 10 μm.
Image from Wang Y et al., PLoS One. 2017;12(7):e0180731. Fig 3.; doi: 10.1371/journal.pone.0180731. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/Immunocytochemistry/ Immunofluorescence - Anti-GRP78 BiP antibody (ab21685)
HEK-293 cells were cultured on coverslips and transiently transfected with wild-type murine IGSF1-2-Myc/His, premeabilized, and processed for double-label immunofluorescence with the Myc antibody (green) and an antibody against GRP78 BiP (red). The overlay is shown in yellow. Nuclei were stained with DAPI. Images were captured by confocal microscopy. Scale bar, 10 μm.
Western blot - Anti-GRP78 BiP antibody (ab21685)
Blocking/Diluting buffer and concentration: 5% NFDM/TBST
All lanes: Western blot - Anti-GRP78 BiP antibody (ab21685) at 1/1000 dilution
Lane 1: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 20 µg
Lane 2: HeLa treated with 2.5 μg/ml tunicamycin for 24h whole cell lysates at 20 µg
Lane 3: HUVEC (Human umbilical vein endothelial cell) whole cell lysates at 20 µg
Lane 4: HUEVC (Human umbilical vein endothelial cell) treated with 10 μg/ml tunicamycin for 48h whole cell lysates at 20 µg
Lane 5: Raw 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysates at 20 µg
Lane 6: Raw 264.7 treated with 5 μg/ml tunicamycin for 18h whole cell lysates at 20 µg
SecondaryAll lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 72 kDa
Observed band size: 78 kDa
Exposure time: 3s
Western blot - Anti-GRP78 BiP antibody (ab21685)
ab21685 recognises a band of ~ 75 kDa in CHO, mouse liver, rat liver and HeLa whole cell lysates, corresponding to GRP78 BiP. This band is quenched by the addition of the immunizing peptide, ab22410.
ab21685 also detects a 100 kDa band in Western Blot. We are unsure of the identity of this protein.
All lanes: Western blot - Anti-GRP78 BiP antibody (ab21685) at 1 µg/mL
Lane 1: CHO-K1 (chinese hamster ovary cell line) whole cell lysate at 20 µg
Lanes 2 and 6: Liver (Mouse) Tissue Lysate at 20 µg
Lanes 3 and 7: Rat liver whole cell lysate at 20 µg
Lane 4: HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 5: CHO-K1 whole cell lysate at 20 µg/mL
Lane 8: HeLa whole cell lysate at 20 µg
SecondaryAll lanes: Goat anti Rabbit IgG at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 72 kDa
Observed band size: 100 kDa, 75 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GRP78 BiP antibody (ab21685)
IHC image of GRP78 BiP staining in human liver carcinoma FFPE section, performed on a BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab21685, 1µg/ml, for 8 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
Western blot - Anti-GRP78 BiP antibody (ab21685)
All lanes: Western blot - Anti-GRP78 BiP antibody (ab21685) at 1 µg/mL
Lane 1: CHO-K1 (chinese hamster ovary cell line) Whole Cell Lysate
Lane 2: Liver (Mouse) Tissue Lysate at 10 µg
Lane 3: Liver (Rat) Tissue Lysate at 10 µg
Lane 4: HeLa (human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg
SecondaryAll lanes: Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 72 kDa
Observed band size: 100 kDa, 78 kDa
Exposure time: 1min
This image is courtesy of an anonymous customer review.Immunocytochemistry/ Immunofluorescence - Anti-GRP78 BiP antibody (ab21685)
Immunocytochemistry/ Immunofluorescence analysis of MDA-MB-435S tumor cell line cells labeling GRP78 BiP with ab21685 at 1/400 dilution. Cells were fixed in formaldehyde and permeabilized with 0.25% Triton-X100 in PBS for 10 minutes. Blocking was done with 1%BSA for 1 hour at 20°C; followed by staining with ab21685 at 1/400 for 18 hours. Undiluted Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077, a Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) secondary antibody was used. DAPI was used to counterstain.
Western blot - Anti-GRP78 BiP antibody (ab21685)
All lanes: Western blot - Anti-GRP78 BiP antibody (ab21685) at 1 µg/mL
Lane 1: Western blot - Recombinant human GRP78 BiP protein (Active) (Recombinant human GRP78 BiP protein (Active) ab78432) at 0.1 µg
Lane 2: Western blot - Recombinant human GRP78 BiP protein (Active) (Recombinant human GRP78 BiP protein (Active) ab78432) at 0.01 µg
SecondaryAll lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (Goat Anti-Rabbit IgG H&L (HRP) preadsorbed ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 72 kDa
Exposure time: 10s
Immunocytochemistry/ Immunofluorescence - Anti-GRP78 BiP antibody (ab21685)
ab21685 staining GRP78 BiP in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab21685 at 1µg/ml and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour magenta). Nuclear DNA was labelled with DAPI (shown in blue).
Also suitable in cells fixed with 4% paraformaldehyde (10 min).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GRP78 BiP antibody (ab21685)
Immunohistochemical analysis of formalin fixed paraffin embedded human liver carcinoma labelling GRP78 BiP with ab21685 at a concentration of 0.4µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins.
ab21685 Anti-GRP78 BiP antibody was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.
Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).
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