Overview
- Product nameAnti-GRP78 BiP antibodySee all GRP78 BiP primary antibodies ...
- DescriptionRabbit polyclonal to GRP78 BiP
- Tested applicationsIP, ICC/IF, WB, Electron Microscopy, IHC-P more details
- Species reactivityReacts 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 controlThis 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
- FormLiquid
- Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
- Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 -
Concentration information loading... - PurityImmunogen affinity purified
- Clonality Polyclonal
- IsotypeIgG
- Research Areas
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 | Notes |
|---|---|
| IP | IP: Use a concentration of 2 µg/ml. |
| ICC/IF | ICC/IF: Use a concentration of 1 µg/ml. |
| WB | WB: Use 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). |
| Electron Microscopy | EM: 1/250. |
| IHC-P | IHC-P: Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
Target
- FunctionProbably plays a role in facilitating the assembly of multimeric protein complexes inside the ER.
- Involvement in diseaseNote=Autoantigen in rheumatoid arthritis.
- Sequence similaritiesBelongs to the heat shock protein 70 family.
- Cellular localizationEndoplasmic reticulum lumen. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
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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
- Unigene: 605502 Human
- Unigene: 743241 Human
- Unigene: 330160 Mouse
- Unigene: 470180 Mouse
- Unigene: 11088 Rat
see all
Target information above from: UniProt accession
P11021
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010)
.
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Alternative names
- 78 kDa glucose regulated protein antibody78 kDa glucose-regulated protein antibodyAL022860 antibody
- AU019543 antibodyBIP antibodyD2Wsu141e antibodyD2Wsu17e antibodyEndoplasmic reticulum lumenal Ca(2+)-binding protein grp78 antibodyEndoplasmic reticulum lumenal Ca2+ binding protein grp78 antibodyFLJ26106 antibodyGlucose Regulated Protein 78kDa antibodyGRP 78 antibodyGRP-78 antibodyGRP78 antibodyGRP78_HUMAN antibodyHeat shock 70 kDa protein 5 antibodyHeat Shock 70kDa Protein 5 antibodyHsce70 antibodyHSPA 5 antibodyHSPA5 antibodyImmunoglobulin Heavy Chain Binding Protein antibodyImmunoglobulin heavy chain-binding protein antibodymBiP antibodyMIF2 antibodySez7 antibody
see all
Anti-GRP78 BiP antibody images
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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 withGRP78 BiP peptide (ab22410) at 1 µg
Lane 6 : Liver (Mouse) Tissue Lysate at 20 µg withGRP78 BiP peptide (ab22410) at 1 µg/ml
Lane 7 : Rat liver whole cell lysate at 20 µg withGRP78 BiP peptide (ab22410) at 1 µg/ml
Lane 8 : HeLa whole cell lysate at 20 µg withGRP78 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. -
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. -
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)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 - 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). -
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 blotAll 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:
- Boyle LH et al. Tapasin-related protein TAPBPR is an additional component of the MHC class I presentation pathway. Proc Natl Acad Sci U S A 110:3465-70 (2013). WB . Read more (PubMed: 23401559) »
- Alapure BV et al. The unfolded protein response is activated in connexin 50 mutant mouse lenses. Exp Eye Res : (2012). Read more (PubMed: 22713599) »
