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Overview
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Product nameAnti-GRP78 BiP antibody
See all GRP78 BiP primary antibodies -
DescriptionRabbit polyclonal to GRP78 BiP
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Host speciesRabbit
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Tested applicationsSuitable for: IHC-FoFr, IP, ICC/IF, WB, Electron Microscopy, IHC-Pmore details
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Species reactivityReacts with: Mouse, Rat, Dog, Human, Pig, African green monkey, Chinese hamster
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Immunogen
Synthetic peptide conjugated to KLH derived from within residues 600 to the C-terminus of Mouse GRP78 BiP.
(Peptide available as ab22410.)
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Positive control
- This antibody gave a positive signal in both Mouse and Rat liver tissue lysates as well as the following whole cell lysates: CHO-K1; HeLa. ICC-IF: Hela cells.
Properties
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FormLiquid
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Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
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Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 -
Concentration information loading... -
PurityImmunogen affinity purified
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ClonalityPolyclonal
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IsotypeIgG
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Research areas
Associated products
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Related Products
Applications
Our Abpromise guarantee covers the use of ab21685 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| IHC-FoFr | Use at an assay dependent concentration. | |
| IP | Use a concentration of 2 µg/ml. | |
| ICC/IF | Use a concentration of 1 - 5 µg/ml. We recommend using Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) secondary antibody. |
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| WB | Use a concentration of 1 µg/ml. Detects a band of approximately 75 kDa (predicted molecular weight: 78 kDa).Can be blocked with Mouse GRP78 BiP peptide (ab22410). | |
| Electron Microscopy | 1/250. | |
| IHC-P | Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
Target
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FunctionProbably plays a role in facilitating the assembly of multimeric protein complexes inside the endoplasmic reticulum. Involved in the correct folding of proteins and degradation of misfolded proteins via its interaction with DNAJC10, probably to facilitate the release of DNAJC10 from its substrate.
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Involvement in diseaseAutoantigen in rheumatoid arthritis.
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Sequence similaritiesBelongs to the heat shock protein 70 family.
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Cellular localizationEndoplasmic reticulum lumen. Melanosome. Cytoplasm. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
- Information by UniProt
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Database links
- Entrez Gene: 100689305 Chinese hamster
- Entrez Gene: 480726 Dog
- Entrez Gene: 3309 Human
- Entrez Gene: 14828 Mouse
- Entrez Gene: 407060 Pig
- Entrez Gene: 25617 Rat
- Omim: 138120 Human
- SwissProt: P11021 Human
see all -
Alternative names
- 78 kDa glucose regulated protein antibody
- 78 kDa glucose-regulated protein antibody
- AL022860 antibody
see all
Images
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Immunocytochemistry/ Immunofluorescence - Anti-GRP78 BiP antibody (ab21685)ab21685 stained in Hela cells. Cells were fixed with 4% paraformaldehyde (10min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% triton for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab21685 at 1µg/ml and ab7291 (Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control) at 1/1000 dilution overnight at +4°C. The secondary antibodies were Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) (pseudo-colored red) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed (ab150081) secondary antibody (colored green) used at 1 ug/ml for 1hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43µM for 1hour at room temperature.
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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 : Anti-GRP78 BiP antibody (ab21685) at 1 µg/ml
Lane 1 : CHO-K1 whole cell lysate at 20 µg
Lane 2 : Liver (Mouse) Tissue Lysate at 20 µg
Lane 3 : Rat liver whole cell lysate at 20 µg
Lane 4 : HeLa whole cell lysate at 20 µg
Lane 5 : CHO-K1 whole cell lysate at 20 µg/ml withMouse GRP78 BiP peptide (ab22410) at 1 µg
Lane 6 : Liver (Mouse) Tissue Lysate at 20 µg withMouse GRP78 BiP peptide (ab22410) at 1 µg/ml
Lane 7 : Rat liver whole cell lysate at 20 µg withMouse GRP78 BiP peptide (ab22410) at 1 µg/ml
Lane 8 : HeLa whole cell lysate at 20 µg withMouse GRP78 BiP peptide (ab22410) at 1 µg/ml
Secondary
All lanes : Goat anti Rabbit IgG at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 78 kDa
Observed band size: 75 kDa (why is the actual band size different from the predicted?)
Additional bands at: 100 kDa. We are unsure as to the identity of these extra bands.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.
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Western blot - Anti-GRP78 BiP antibody (ab21685)All lanes : Anti-GRP78 BiP antibody (ab21685) at 1 µg/ml
Lane 1 : CHO-K1 cell lysate 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
Secondary
All 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: 78 kDa
Observed band size: 78 kDa
Additional bands at: 100 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 1 minute -
Immunocytochemistry/ Immunofluorescence - Anti-GRP78 BiP antibody (ab21685)This image is courtesy of an anonymous Abreviewab21685 staining GRP78 BiP in African Green Monkey COS-7 cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.5% Triton X-100 and blocked with 3% BSA for 1 hour at 23°C. Samples were incubated with primary antibody (1/1000 in PBS-BSA) for 1 hour at 23°C. An Alexa Fluor® 488-conjugated Goat anti-rabbit IgG polyclonal (1/1000) was used as the secondary antibody.
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Immunocytochemistry/ Immunofluorescence - Anti-GRP78 BiP antibody (ab21685)This image was kindly supplied by Dr Vladimir Milenkovic by Abreviewab21685 at a 1/500 dilution staining GRP78 BiP in mouse retinal pigment epithelium primary cells by Immunocytochemistry/ Immunofluorescence, incubated for 16 hours at 4°C. PFA fixed. Blocked with 5% serum for 20 minutes at 25°C. Secondary used at a 1/500 dilution polyclonal Goat anti-rabbit conjugated to Alexa Fluor 488 (green). Nuclei were counterstained with DAPI (blue). -
Immunocytochemistry/ Immunofluorescence - Anti-GRP78 BiP antibody (ab21685)This image was kindly supplied by Dr Vladimir Milenkovic by Abreviewab21685 at a 1/500 dilution staining GRP78 BiP in Dog MDCK II cells by Immunocytochemistry/ Immunofluorescence, incubated for 16 hours at 4°C. PFA fixed. Blocked with 5% serum for 20 minutes at 25°C. Secondary used at a 1/500 dilution polyclonal Goat anti-rabbit conjugated to Alexa Fluor 488 (green). Nuclei were counterstained with DAPI (blue). -
Western blot - Anti-GRP78 BiP antibody (ab21685)All lanes : Anti-GRP78 BiP antibody (ab21685) at 1 µg/ml
Lane 1 :Recombinant human GRP78 BiP protein (His tag) (ab78432) at 0.1 µg
Lane 2 :Recombinant human GRP78 BiP protein (His tag) (ab78432) at 0.01 µg
Secondary
All lanes : 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: 78 kDa
Exposure time: 10 seconds -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GRP78 BiP antibody (ab21685)This image is courtesy of an anonymous abreview.ab21685 staining Panc1 tumor taken out of Balb/C nude mice sections by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). The tissue was fixed with formaldehyde and a heat mediated antigen retrival step was performed with citric acid pH 6. Staining with ab21685 at 1 µg/ml for 18 hours at 4°C. An undiluted goat anti-rabbit polyclonal HRP conjugated secondary antibody was used.
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Immunocytochemistry/ Immunofluorescence - Anti-GRP78 BiP antibody (ab21685)This image is courtesy of an anonymous abreview.Immunocytochemistry/ Immunofluorescence analysis of MDA-MB-435 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 ab150077, a Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) secondary antibody was used. DAPI was used to counterstain.
Protocols
References
This product has been referenced in:
- Zheng WL et al. ROS-Mediated Cell Cycle Arrest and Apoptosis Induced by Zearalenone in Mouse Sertoli Cells via ER Stress and the ATP/AMPK Pathway. Toxins (Basel) 10:N/A (2018). Read more (PubMed: 29301253) »
- Leipnitz G et al. Evaluation of mitochondrial bioenergetics, dynamics, endoplasmic reticulum-mitochondria crosstalk, and reactive oxygen species in fibroblasts from patients with complex I deficiency. Sci Rep 8:1165 (2018). WB ; Human . Read more (PubMed: 29348607) »
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