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, Flow Cyt, IPmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human GRP94 aa 100-200. The exact sequence is proprietary.
    Database link: P14625

  • Positive control

    • WB: Human kidney and stomach tissue lysates; mouse brain and placenta tissue lysates; rat brain, placenta and stomach tissue lysates; HeLa, MCF7, 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: HeLa cells. IP: HeLa whole cell lysate.
  • General notes

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

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.
Flow Cyt 1/600.
IP 1/30.

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.
  • Information by UniProt
  • Database links

  • 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

  • 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.

  • 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.

  • 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.

  • 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 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.

  • 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).

  • 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).

     

  • 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.

     

  • 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.

     

References

ab238126 has not yet been referenced specifically in any publications.

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