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  1. Link

    grp94-antibody-epr22847-50-ab238126.pdf

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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-GRP94 antibody [EPR22847-50] (ab238126)

  • Datasheet
  • SDS
  • Certificate of Compliance
Submit a review Submit a question References (4)

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Western blot - Anti-GRP94 antibody [EPR22847-50] (ab238126)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GRP94 antibody [EPR22847-50] (ab238126)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GRP94 antibody [EPR22847-50] (ab238126)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GRP94 antibody [EPR22847-50] (ab238126)
  • Immunoprecipitation - Anti-GRP94 antibody [EPR22847-50] (ab238126)
  • Flow Cytometry (Intracellular) - Anti-GRP94 antibody [EPR22847-50] (ab238126)
  • Western blot - Anti-GRP94 antibody [EPR22847-50] (ab238126)
  • Western blot - Anti-GRP94 antibody [EPR22847-50] (ab238126)
  • Immunocytochemistry/ Immunofluorescence - Anti-GRP94 antibody [EPR22847-50] (ab238126)
  • Immunocytochemistry/ Immunofluorescence - Anti-GRP94 antibody [EPR22847-50] (ab238126)
  • Anti-GRP94 antibody [EPR22847-50] (ab238126)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR22847-50] to GRP94
  • Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra)
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

You may also be interested in

Secondary
Product image
Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
Knockout
Product image
Human HSP90B1 (GRP94) knockout HEK-293T cell line (ab266313)
Protein
Product image
Recombinant Human GRP94 protein (ab87144)

View more associated products

Overview

  • Product name

    Anti-GRP94 antibody [EPR22847-50]
    See all GRP94 primary antibodies
  • Description

    Rabbit monoclonal [EPR22847-50] to GRP94
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cyt (Intra)more details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Human kidney and stomach tissue lysates; mouse brain and placenta tissue lysates; rat brain, placenta and stomach tissue lysates; HeLa, MCF7, HEK293T, MEF, PC-12, C6 and NIH/3T3 whole cell lysates. IHC-P: Human kidney tissue; mouse and rat colon tissue. ICC/IF: HeLa and NIH/3T3 cells. Flow Cyt (intra): HeLa cells. IP: HeLa whole cell lysate.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: PBS, 0.05% BSA, 40% Glycerol (glycerin, glycerine)
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR22847-50
  • Isotype

    IgG
  • Research areas

    • Tags & Cell Markers
    • Subcellular Markers
    • Organelles
    • ER
    • Signal Transduction
    • Protein Trafficking
    • ER Proteins
    • Cancer
    • Cancer Metabolism
    • Response to hypoxia
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Hypoxia

Associated products

  • Alternative Versions

    • Anti-GRP94 antibody [EPR22847-50] - BSA and Azide free (ab256312)
  • Compatible Secondaries

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)
    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Immunohistochemistry kits

    • Rabbit specific IHC polymer detection kit HRP/DAB (ab209101)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human HSP90B1 (GRP94) knockout HEK-293T cell line (ab266313)
  • KO cell lysates

    • Human HSP90B1 (GRP94) knockout HEK-293T cell lysate (ab257254)
  • Recombinant Protein

    • Recombinant Human GRP94 protein (ab87144)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab238126 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
WB
1/1000. Detects a band of approximately 94 kDa (predicted molecular weight: 94 kDa).
IHC-P
1/5000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
1/1000.
IP
1/30.
Flow Cyt (Intra)
1/600.
Notes
WB
1/1000. Detects a band of approximately 94 kDa (predicted molecular weight: 94 kDa).
IHC-P
1/5000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
1/1000.
IP
1/30.
Flow Cyt (Intra)
1/600.

Target

  • Function

    Molecular chaperone that functions in the processing and transport of secreted proteins. Functions in endoplasmic reticulum associated degradation (ERAD). Has ATPase activity.
  • Sequence similarities

    Belongs to the heat shock protein 90 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 accession P14625 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 7184 Human
    • Entrez Gene: 22027 Mouse
    • Entrez Gene: 362862 Rat
    • Omim: 191175 Human
    • SwissProt: P14625 Human
    • SwissProt: P08113 Mouse
    • SwissProt: Q66HD0 Rat
    • Unigene: 192374 Human
    • Unigene: 87773 Mouse
    • Unigene: 101146 Rat
    see all
  • Alternative names

    • 94 kDa glucose regulated protein antibody
    • 94 kDa glucose-regulated protein antibody
    • ECGP antibody
    • Endoplasmin antibody
    • Endothelial cell (HBMEC) glycoprotein antibody
    • ENPL_HUMAN antibody
    • Glucose regulated protein 94kDa antibody
    • gp96 antibody
    • gp96 homolog antibody
    • GRP 94 antibody
    • GRP-94 antibody
    • Heat shock protein 90 kDa beta member 1 antibody
    • heat shock protein 90kDa beta (Grp94), member 1 antibody
    • Heat shock protein, 90 kDa, beta, 1 antibody
    • HSP90B1 antibody
    • Stress inducible tumor rejection antigen GP96 antibody
    • TRA1 antibody
    • tumor rejection antigen (gp96) 1 antibody
    • Tumor rejection antigen 1 antibody
    • Tumor rejection antigen gp96 antibody
    • Tumor rejection antigen-1 (gp96) antibody
    see all

Images

  • Western blot - Anti-GRP94 antibody [EPR22847-50] (ab238126)
    Western blot - Anti-GRP94 antibody [EPR22847-50] (ab238126)
    All lanes : Anti-GRP94 antibody [EPR22847-50] (ab238126) at 1/1000 dilution

    Lane 1 : Wild-type HEK293T cell lysate
    Lane 2 : HSP90B1 knockout HEK293T cell lysate
    Lane 3 : HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 94 kDa
    Observed band size: 94 kDa



    Lanes 1-3: Merged signal (red and green). Green - ab238126 observed at 94 kDa. Red - loading control, ab8245 observed at 37 kDa.

    ab238126 Anti-GRP94 antibody [EPR22847-50] was shown to specifically react with GRP94 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266313 (knockout cell lysate ab257254) was used. Wild-type and GRP94 knockout samples were subjected to SDS-PAGE. ab238126 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GRP94 antibody [EPR22847-50] (ab238126)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GRP94 antibody [EPR22847-50] (ab238126)

    Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling GRP94 with ab238126 at 1/5000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on rat colon (PMID: 23572575) is observed. Counterstained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    The section was incubated with ab238126 for 15 mins at RT.

    The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GRP94 antibody [EPR22847-50] (ab238126)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GRP94 antibody [EPR22847-50] (ab238126)

    Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling GRP94 with ab238126 at 1/5000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on mouse colon (PMID: 23572575) is observed. Counterstained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    The section was incubated with ab238126 for 15 mins at RT.

    The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GRP94 antibody [EPR22847-50] (ab238126)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GRP94 antibody [EPR22847-50] (ab238126)

    Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling GRP94 with ab238126 at 1/5000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining on human kidney (PMID: 20520781) is observed. Counterstained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    The section was incubated with ab238126 for 15 mins at RT.

    The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

  • Immunoprecipitation - Anti-GRP94 antibody [EPR22847-50] (ab238126)
    Immunoprecipitation - Anti-GRP94 antibody [EPR22847-50] (ab238126)

    GRP94 was immunoprecipitated from 0.35 mg of HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab238126 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab238126 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used at 1/5000 dilution.

    Lane 1: HeLa whole cell lysate 10 μg (Input).
    Lane 2: ab238126 IP in HeLa whole cell lysate.
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab238126 in HeLa whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.
    Exposure time: 3 seconds.

  • Flow Cytometry (Intracellular) - Anti-GRP94 antibody [EPR22847-50] (ab238126)
    Flow Cytometry (Intracellular) - Anti-GRP94 antibody [EPR22847-50] (ab238126)

    Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling GRP94 with ab238126 at 1/600 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). 

    Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077), at 1/2000 dilution was used as the secondary antibody. 

     

     

  • Western blot - Anti-GRP94 antibody [EPR22847-50] (ab238126)
    Western blot - Anti-GRP94 antibody [EPR22847-50] (ab238126)
    All lanes : Anti-GRP94 antibody [EPR22847-50] (ab238126) at 1/1000 dilution

    Lane 1 : Mouse brain tissue lysate at 20 µg
    Lane 2 : Mouse placenta tissue lysate at 20 µg
    Lane 3 : Mouse stomach tissue lysate at 20 µg
    Lane 4 : Rat brain tissue lysate at 20 µg
    Lane 5 : Rat placenta tissue lysate at 20 µg
    Lane 6 : Rat stomach tissue lysate at 20 µg
    Lane 7 : C6 (rat glial tumor glial cell line) whole cell lysate at 20 µg
    Lane 8 : NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate at 10 µg

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 94 kDa
    Observed band size: 94 kDa



    Blocking and dilution buffer: 5% NFDM/TBST.

    Exposure times.

    Lane 1: 26 seconds;  Lanes 2-5: 10 seconds; Lane 6: 92 seconds; Lanes 7-8: 6 seconds.

    The molecular weight observed is consistent with what has been described in the literature (PMID: 26858453, 29949743).

  • Western blot - Anti-GRP94 antibody [EPR22847-50] (ab238126)
    Western blot - Anti-GRP94 antibody [EPR22847-50] (ab238126)
    All lanes : Anti-GRP94 antibody [EPR22847-50] (ab238126) at 1/1000 dilution

    Lane 1 : Human brain tissue lysate at 20 µg
    Lane 2 : Human kidney tissue lysate at 20 µg
    Lane 3 : Human stomach tissue lysate at 20 µg
    Lane 4 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
    Lane 5 : MCF7 (human breast adenocarcinoma cell line) whole cell lysate at 20 µg
    Lane 6 : MEF (Mouse embryonic fibroblast (immortalized) cell line) whole cell lysate at 20 µg
    Lane 7 : PC-12 (rat adrenal gland pheochromocytoma cell line) whole cell lysate at 10 µg

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 94 kDa
    Observed band size: 94 kDa



    Blocking and dilution buffer: 5% NFDM/TBST.

    Exposure times.

    Lanes 1 & 2: 10 seconds; Lane 3: 37 seconds; Lanes 4-6: 10 seconds; Lane 7: 26 seconds.

    The molecular weight observed is consistent with what has been described in the literature (PMID: 26858453, 29949743).

     

  • Immunocytochemistry/ Immunofluorescence - Anti-GRP94 antibody [EPR22847-50] (ab238126)
    Immunocytochemistry/ Immunofluorescence - Anti-GRP94 antibody [EPR22847-50] (ab238126)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (mouse embryo fibroblast cell line) cells labeling GRP94 with ab238126 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000  dilution (green). Confocal image showing endoplasmic reticulum staining in NIH/3T3 cell line. The nuclear counterstain is DAPI (blue). The endoplasmic reticulum is stained with Anti-KDEL antibody [10C3] (ab12223), followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) secondary antibody (red).

    Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000  dilution.

     

  • Immunocytochemistry/ Immunofluorescence - Anti-GRP94 antibody [EPR22847-50] (ab238126)
    Immunocytochemistry/ Immunofluorescence - Anti-GRP94 antibody [EPR22847-50] (ab238126)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling GRP94 with ab238126 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000  dilution (green). Confocal image showing endoplasmic reticulum staining in HeLa cell line. The nuclear counter stain is DAPI (blue). The endoplasmic reticulum is stained with Anti-KDEL antibody [10C3] (ab12223), followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed (ab150120) secondary antibody (red).

    Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000  dilution.

     

  • Anti-GRP94 antibody [EPR22847-50] (ab238126)
    Anti-GRP94 antibody [EPR22847-50] (ab238126)

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

Certificate of Compliance

To download a Certificate of Compliance, please enter your Lot number below:

References (4)

Publishing research using ab238126? Please let us know so that we can cite the reference in this datasheet.

ab238126 has been referenced in 4 publications.

  • Zhang Y  et al. Phosphatase Shp2 regulates biogenesis of small extracellular vesicles by dephosphorylating Syntenin. J Extracell Vesicles 10:e12078 (2021). PubMed: 33732417
  • Persaud AK  et al. Facilitative lysosomal transport of bile acids alleviates ER stress in mouse hematopoietic precursors. Nat Commun 12:1248 (2021). PubMed: 33623001
  • Mo C  et al. LncRNA nuclear-enriched abundant transcript 1 shuttled by prostate cancer cells-secreted exosomes initiates osteoblastic phenotypes in the bone metastatic microenvironment via miR-205-5p/runt-related transcription factor 2/splicing factor proline- and glutamine-rich/polypyrimidine tract-binding protein 2 axis. Clin Transl Med 11:e493 (2021). PubMed: 34459124
  • Jiang K  et al. Exosome-derived ENO1 regulates integrin a6ß4 expression and promotes hepatocellular carcinoma growth and metastasis. Cell Death Dis 11:972 (2020). PubMed: 33184263

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