Astana Biomed Group, an authorized Abcam distributor in Central Asia

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

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 - BSA and Azide free
  • Suitable for: Flow Cyt, ICC/IF, IP, WB, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-GRP94 antibody [EPR22847-50] - BSA and Azide free
    See all GRP94 primary antibodies

  • Description

    Rabbit monoclonal [EPR22847-50] to GRP94 - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Human
    IP
    Human
    WB
    Human
    See all applications and species data

  • 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, HEK293T, 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

    Ab256312 is the carrier-free version of ab238126. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab256312 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

Images

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



    This data was developed using the same antibody clone in a different buffer formulation (ab238126).

    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] - BSA and Azide free (ab256312)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GRP94 antibody [EPR22847-50] - BSA and Azide free (ab256312)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab238126).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GRP94 antibody [EPR22847-50] - BSA and Azide free (ab256312)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GRP94 antibody [EPR22847-50] - BSA and Azide free (ab256312)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab238126).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GRP94 antibody [EPR22847-50] - BSA and Azide free (ab256312)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GRP94 antibody [EPR22847-50] - BSA and Azide free (ab256312)

    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. Counter stained 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab238126).
  • Immunoprecipitation - Anti-GRP94 antibody [EPR22847-50] - BSA and Azide free (ab256312)
    Immunoprecipitation - Anti-GRP94 antibody [EPR22847-50] - BSA and Azide free (ab256312)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab238126).

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

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab238126).

  • Immunocytochemistry/ Immunofluorescence - Anti-GRP94 antibody [EPR22847-50] - BSA and Azide free (ab256312)
    Immunocytochemistry/ Immunofluorescence - Anti-GRP94 antibody [EPR22847-50] - BSA and Azide free (ab256312)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab238126).

  • Immunocytochemistry/ Immunofluorescence - Anti-GRP94 antibody [EPR22847-50] - BSA and Azide free (ab256312)
    Immunocytochemistry/ Immunofluorescence - Anti-GRP94 antibody [EPR22847-50] - BSA and Azide free (ab256312)

    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 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab238126).

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

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