Overview

  • Product nameAnti-GRP94 antibody
    See all GRP94 primary antibodies
  • Description
    Rabbit polyclonal to GRP94
  • SpecificityAt a very high concentration (<1/100), this antibody may recognise PDI and calreticulin. We recommend using this antibody at a relatively high dilution.
  • Tested applicationsSuitable for: ICC, WB, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Whole protein from dog pancreas.

Properties

Applications

Our Abpromise guarantee covers the use of ab3674 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
ICC 1/100.
WB 1/2000 - 1/5000. Detects a band of approximately 94 kDa. Can be used at dilutions up to 1/30,000.
IHC-P 1/1000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF Use at an assay dependent concentration. Used at a concentration of 1/100 for 30 min on human renal carcinoma cell (see Abreview).

Target

  • FunctionMolecular chaperone that functions in the processing and transport of secreted proteins. Functions in endoplasmic reticulum associated degradation (ERAD). Has ATPase activity.
  • Sequence similaritiesBelongs to the heat shock protein 90 family.
  • Cellular localizationEndoplasmic 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

Anti-GRP94 antibody images

  • Immunostaining of mouse embryonic fibroblasts with ab3674. (See Nakamura et al. 2001).
  • Anti-GRP94 antibody (ab3674) at 1/2000 dilution + Cell lysates prepared from mouse fibroblasts
  • ab3674 at a 1/100 dilution staining human renal cell carcinoma cells by ICC/IF. The cells were permeabilized with 0.2% triton X100 and then blocked with 1%BSA, 4% goat serum prior to incubation with the primary 1antibody for 30 min at 37 °C. Bound antibody was detected using an Alexa fluor ® 488 Goat anti-Rabbit IgG (H+L).

    See Abreview

  • Ab3674 (1/1000) staining GPR94 in human stomach using an automated system (DAKO Autostainer Plus). Using this protocol there is strong cytoplasmic staining of peptic and parietal cells of the gastric gland.
    Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.

  • All lanes : Anti-GRP94 antibody (ab3674) at 1/2000 dilution

    Lane 1 : Whole cell lysate prepared from murine 3T3 cells
    Lane 2 : Whole cell lysate prepared from murine 3T3 cells, treated with 2.5 ug/ml Tunicamycin 10 hours

    Lysates/proteins at 50 µg per lane.

    Secondary
    HRP conjugated goat anti-rabbit IgG at 1/2000 dilution
    developed using the ECL technique

    Observed band size : 94 kDa (why is the actual band size different from the predicted?)


    Exposure time : 30 seconds

    Image courtesy of an anonymous Abreview.

    See Abreview

References for Anti-GRP94 antibody (ab3674)

This product has been referenced in:

See all 7 Publications for this product

Product Wall

Application Western blot
Loading amount 25 µg
Gel Running Conditions Reduced Denaturing
Sample Mouse Cell lysate - whole cell (hepatocytes)
Specification hepatocytes
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
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Submitted Dec 25 2014

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Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (Bonemarrow)
Specification Bonemarrow
Fixative Paraformaldehyde
Permeabilization No
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Submitted Apr 25 2012

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Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Fibroblast)
Specification Fibroblast
Fixative Formaldehyde
Permeabilization Yes - 0.1 Triton X-100
Blocking step BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
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Submitted Nov 23 2011

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Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (3T3)
Specification 3T3
Fixative Formaldehyde
Permeabilization Yes - Saponin
Blocking step BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
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Submitted Oct 31 2011

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Application Western blot
Sample Human Cell lysate - whole cell (Macrophages)
Loading amount 40 µg
Specification Macrophages
Treatment Thapsigargin for 24 hours
Gel Running Conditions Reduced Denaturing (12%)
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
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Submitted Sep 30 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - whole cell (3T3)
Loading amount 50 µg
Specification 3T3
Treatment 2.5 ug/ml Tunicamycin 10 hours
Gel Running Conditions Reduced Denaturing (10%)
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
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Submitted Jul 28 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HEK293 EBNA)
Specification HEK293 EBNA
Fixative 3% Paraformaldehyde
Permeabilization No
Blocking step NH4Cl as blocking agent for 15 minute(s) · Concentration: 0.27% · Temperature: 20°C
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Submitted Mar 22 2010

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Application Western blot
Sample Mouse Cell lysate - whole cell (lymphnode and DCEK mouse fibroblast cell)
Loading amount 30 µg
Specification lymphnode and DCEK mouse fibroblast cell
Gel Running Conditions Reduced Denaturing (10%)
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted Aug 20 2009

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Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Fixative Paraformaldehyde
Permeabilization Yes - 0.1% Triton X-100 (10 mins)
Blocking step BSA as blocking agent for 15 minute(s) · Concentration: 3% · Temperature: RT°C
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Dr. Dan Tattersall

Verified customer

Submitted Nov 10 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Renal Carcinoma Cell Line)
Specification Renal Carcinoma Cell Line
Fixative Paraformaldehyde
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 4%
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Submitted Jul 12 2006

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