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Products:Tags & Cell Markers >> Subcellular Markers >> Organelles >> ER
Overview
Product name
Anti-GRP78 BiP antibody
See all GRP78 BiP products (13) ...
Description
Rabbit polyclonal to GRP78 BiP
Tested applications
IP, ICC/IF, WB, Electron Microscopy, IHC-Pmore details
Cross reactivity
Reacts with
Mouse, Rat, Dog, Human, Pig, Chinese Hamster
Immunogen
Synthetic peptide conjugated to KLH derived from within residues 600 to the C-terminus of Mouse GRP78 BiP.
(Peptide available as ab22410.)
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.
Properties
Form
Liquid
Storage instructions
Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage buffer
Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration
Concentration information loading...
Purity
Immunogen affinity purified
Clonality
Polyclonal
Isotype
IgG
Research areas
Tags & Cell Markers >> Subcellular Markers >> Organelles >> ER
Applications
Show applications key
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.
Electron Microscopy
Application notes
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.
Target
Function
Probably plays a role in facilitating the assembly of multimeric protein complexes inside the ER.
Involvement in disease
Note=Autoantigen in rheumatoid arthritis.
Sequence similarities
Belongs to the heat shock protein 70 family.
Cellular localization
Endoplasmic reticulum lumen. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
Target information above from: UniProt accessionP11021
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).
Alternative names
- 78 kDa glucose regulated protein antibody
- 78 kDa glucose-regulated protein antibody
- BIP antibody
see all
Database links
- Entrez Gene: 480726 Dog
- Entrez Gene: 3309 Human
- Entrez Gene: 14828 Mouse
- Entrez Gene: 25617 Rat
- Omim: 138120 Human
- SwissProt: P11021 Human
- SwissProt: P20029 Mouse
- SwissProt: P06761 Rat
see all
Anti-GRP78 BiP antibody images:
Western blot - 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 with GRP78 BiP peptide (ab22410) at 1 µg
Lane 6 : Liver (Mouse) Tissue Lysate at 20 µg 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.
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).
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).
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.
Western blot - 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
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 - GRP78 BiP antibody (ab21685)
ab21685 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).
This image was kindly supplied by Dr Vladimir Milenkovic by Abreview
Immunocytochemistry/ Immunofluorescence - GRP78 BiP antibody (ab21685)
ab21685 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).
This image was kindly supplied by Dr Vladimir Milenkovic by Abreview
Immunocytochemistry/ Immunofluorescence - GRP78 BiP antibody (ab21685)
ICC/IF image of ab21685 stained Hela cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab21685, 1µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-rabbit IgG - H&L, pre-adsorbed (ab96899) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Western blot
All lanes : Anti-GRP78 BiP antibody (ab21685) at 1 µg/ml
Lane 1 : GRP78 BiP protein (Active) (ab78432) at 0.1 µg
Lane 2 : GRP78 BiP protein (Active) (ab78432) at 0.01 µg
Secondary
Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed (ab97080) at 1/5000 dilution
developed using the ECL technique
Performed under reducing conditions.
Exposure time : 10 seconds
References for Anti-GRP78 BiP antibody (ab21685)
This product has been referenced in:
- Randazzo Jet al. Orally active multi-functional antioxidants delay cataract formation in streptozotocin (type 1) diabetic and gamma-irradiated rats. PLoS One 6:e18980 (2011). WB; Rat.Read more (PubMed: 21541328) »
- Das S & Pellett PE Spatial Relationships between Markers for Secretory and Endosomal Machinery in Human Cytomegalovirus-Infected Cells versus Those in Uninfected Cells. J Virol 85:5864-79 (2011).Read more (PubMed: 21471245) »
See all 22 publications for this product
Publishing research using ab21685? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"