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Western blot - GRP78 BiP antibody (ab21685)
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Immunocytochemistry/ Immunofluorescence - GRP78 BiP antibody (ab21685)
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Immunocytochemistry/ Immunofluorescence - GRP78 BiP antibody (ab21685)
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Product Name
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GRP78 BiP antibody
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See all GRP78 BiP - ER marker antibodies (6)...
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Product type
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Primary antibodies
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Description
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Rabbit polyclonal to GRP78 BiP
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Immunogen
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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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Reacts with
(species key)
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Hu, Ms, Rat, ChHm, Dog, Pig
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Tested applications
(see key)
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EM, ICC/IF, IHC-P, IP, WB
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Abreviews
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Application notes
(see key)
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Recommended dilutions
EM: 1/250 (PMID 19209902).
ICC/IF: Use at a concentration of 1 µg/ml.
IHC-P: Use at a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IP: Use at a concentration of 2 µg/ml.
WB: Use at a concentration of 1 µg/ml. Detects a band of approximately 75 kDa (predicted molecular weight: 78 kDa). Can be blocked with GRP78 BiP peptide (ab22410).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
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Positive control
(see definition)
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CHO K1 cell lysate. Mouse and Rat liver cell lysate. HeLa whole cell lysate
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Cellular localization
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Endoplasmic reticulum lumen.
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Research areas
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Tags & Cell Markers >> Subcellular Markers >> Organelles >> ER
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Relevance
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The 78 kDa glucose regulated protein/BiP (GRP78) belongs to the family of ~70 kDa heat shock proteins (HSP 70). GRP78 is a resident protein of the endoplasmic reticulum (ER) and may associate transiently with a variety of newly synthesized secretory and membrane proteins or permanently with mutant or defective proteins that are incorrectly folded, thus preventing their export from the ER lumen. GRP78 is a highly conserved protein that is essential for cell viability. The highly conserved sequence Lys-Asp-Glu-Leu (KDEL) is present at the C terminus of GRP78 and other resident ER proteins including glucose regulated protein 94 (GRP 94) and protein disulfide isomerase (PDI). The presence of carboxy terminal KDEL appears to be necessary for retention and appears to be sufficient to reduce the secretion of proteins from the ER. This retention is reported to be mediated by a KDEL receptor.
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Database links
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The links below go to external sites and will open in a new browser window |
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Raised in
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Rabbit
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Clonality
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Polyclonal
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Isotype
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IgG
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Purity
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Immunogen affinity purified
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Storage buffer
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Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 Material safety datasheet (MSDS) for this product:
Sodium Azide MSDS
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Form
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Liquid
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Concentration
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Batch dependent within range: 0.10 - 0.30 mg/ml
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Storage instructions
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Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
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At Abcam, we have one centralized database to hold all of our product information, so that everything we know about this GRP78 BiP antibody is on this datasheet. But please do contact us if you would like any reassurance! |
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See below for GRP78 BiP antibody images, references, products related to ab21685 and other tools.
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GRP78 BiP antibody images:
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 Western blot - GRP78 BiP antibody (ab21685)
All lanes : GRP78 BiP antibody (ab21685) at 1 µg/ml
Lane 1 : CHO-K1 whole cell lysate at 20 µg/ml
Lane 2 : Liver (Mouse) Tissue Lysate (ab7935) at 20 µg/ml
Lane 3 : Rat liver whole cell lysate at 20 µg/ml
Lane 4 : HeLa whole cell lysate at 20 µg/ml
Lane 5 : CHO-K1 whole cell lysate at 20 µg/ml with GRP78 BiP peptide (ab22410) at 1 µg/ml
Lane 6 : Liver (Mouse) Tissue Lysate (ab7935) at 20 µg/ml with GRP78 BiP peptide (ab22410) at 1 µg/ml
Lane 7 : Rat liver whole cell lysate at 20 µg with GRP78 BiP peptide (ab22410) at 1 µg/ml
Lane 8 : HeLa whole cell lysate at 20 µg with GRP78 BiP peptide (ab22410) at 1 µg/ml
Secondary
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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Immunocytochemistry/ Immunofluorescence - GRP78 BiP antibody (ab21685)
ICC/IF image of ab21685 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab21685, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).
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Immunocytochemistry/ Immunofluorescence - GRP78 BiP antibody (ab21685)
ICC/IF image of ab21685 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab21685, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - 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.
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References for GRP78 BiP antibody (ab21685)
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This product has been used in: (two most recent references)
Zwang NA et al. Identification of phosphorylation-dependent binding partners of aquaporin-2 using protein mass spectrometry. J Proteome Res 8:1540-54 (2009). IHC-P, EM; Rat. PubMed: 19209902
Pasquariello N et al. Characterization of the Endocannabinoid System in Human Neuronal Cells and Proteomic Analysis of Anandamide-induced Apoptosis. J Biol Chem 284:29413-26 (2009). WB; Human. PubMed: 19690173
See all 10 publication references for this product.
If you publish research using ab21685 please let us know so that we can cite the reference on this datasheet.
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Search PubMed (MEDLINE) for references to GRP78 BiP
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GRP78 BiP antibody - more information
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