Overview

  • Product nameAnti-GRP78 BiP antibodySee all GRP78 BiP primary antibodies ...
  • Description
    Rabbit polyclonal to GRP78 BiP
  • Tested applicationsIP, ICC/IF, WB, Electron Microscopy, IHC-P more details
  • Species reactivity
    Reacts with: Mouse, Rat, Dog, Human, Pig, African Green Monkey, 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

  • 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 Use a concentration of 2 µg/ml.
ICC/IF Use a concentration of 1 µg/ml.
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

  • 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.
  • Target information above from: UniProt accession P11021 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links
  • Alternative names
    • 78 kDa glucose regulated protein antibody
    • 78 kDa glucose-regulated protein antibody
    • AL022860 antibody
    • AU019543 antibody
    • BIP antibody
    • D2Wsu141e antibody
    • D2Wsu17e antibody
    • Endoplasmic reticulum lumenal Ca(2+)-binding protein grp78 antibody
    • Endoplasmic reticulum lumenal Ca2+ binding protein grp78 antibody
    • FLJ26106 antibody
    • Glucose Regulated Protein 78kDa antibody
    • GRP 78 antibody
    • GRP-78 antibody
    • GRP78 antibody
    • GRP78_HUMAN antibody
    • Heat shock 70 kDa protein 5 antibody
    • Heat Shock 70kDa Protein 5 antibody
    • Hsce70 antibody
    • HSPA 5 antibody
    • HSPA5 antibody
    • Immunoglobulin Heavy Chain Binding Protein antibody
    • Immunoglobulin heavy chain-binding protein antibody
    • mBiP antibody
    • MIF2 antibody
    • Sez7 antibody
    see all

Anti-GRP78 BiP antibody images

  • 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 Mouse GRP78 BiP peptide (ab22410) at 1 µg
    Lane 6 : Liver (Mouse) Tissue Lysate at 20 µg with Mouse GRP78 BiP peptide (ab22410) at 1 µg/ml
    Lane 7 : Rat liver whole cell lysate at 20 µg with Mouse GRP78 BiP peptide (ab22410) at 1 µg/ml
    Lane 8 : HeLa whole cell lysate at 20 µg with Mouse 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.

    See Abreview

  • 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).

  • 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).

  • 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.
  • 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
  • ab21685 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.

    See Abreview

  • 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).

    See Abreview

  • 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).

    See Abreview

  • 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.

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) »
  • Okamoto Y  et al. Role of C-terminal membrane-proximal basic residues in cell surface trafficking of HIV coreceptor GPR15 protein. J Biol Chem 288:9189-99 (2013). ICC/IF ; Human . Read more (PubMed: 23430259) »

See all 32 Publications for this product

Product Wall

Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 23°C
Sample African Green Monkey Cell (COS-7)
Specification COS-7
Permeabilization Yes - 0.5% Triton-X
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Oct 25 2013

Application Immunoprecipitation
Immuno-precipitation step Protein A
Sample Human Cell lysate - whole cell (prostate cancer cell 22Rv1)
Specification prostate cancer cell 22Rv1
Total protein in input 20 µg
Username

Abcam user community

Verified customer

Submitted Aug 20 2013

Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 4%
Sample Human Cell (prostate cancer cell 22Rv1)
Specification prostate cancer cell 22Rv1
Permeabilization Yes - Triton-X-100
Fixative Paraformaldehyde
Username

Dr. Danilo Maddalo

Verified customer

Submitted Aug 20 2013

Thank you for contacting us.

ab2185 Immunogen sequence is more than 90% similar with Pig HIF1 alpha so this antibody is more likely to cross react.

http://www.ebi.ac.uk/Tools/services/web_clustalw2/toolresult.ebi?tool=clustalw2&jo...

Read More
Application Western blot
Sample Human Cell lysate - whole cell (Breast)
Loading amount 20 µg
Specification Breast
Gel Running Conditions Non-reduced Denaturing
Blocking step Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Abcam user community

Verified customer

Submitted Mar 13 2013

Thank you for contacting us.

For ab21685 we used human liver carcinoma. I would suggest using the following product as a positive control:
Liver tumor (human): hepatocellular carcinoma tissue slides (ab4877) (http://www.abcam.com/ab487...

Read More

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has cau...

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Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (mouse brain)
Specification mouse brain
Fixative Paraformaldehyde
Antigen retrieval step None
Permeabilization No
Blocking step PBS, 10%Horse serum, 0,2%BSA, 0,3%Triton as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 23°C
Username

Abcam user community

Verified customer

Submitted Aug 21 2012

Thank you for your enquiry.

The concentration for this product is lot dependent. We currently have a few concentrations from different lots. Please see below for more details.






1) 1mg/ml (volume 100ul) - L...

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I'm happy to help! Please let me know if you have any further questions.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"