Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [3]
- Immunocytochemistry [1]
- Immunohistochemistry [2]
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Validation data
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- Product number
- GTX22791 - Provider product page
- Provider
- GeneTex
- Proper citation
- GeneTex Cat#GTX22791, RRID:AB_370802
- Product name
- GRP94 antibody [9G10]
- Antibody type
- Monoclonal
- Reactivity
- Human, Mouse, Rat, Bovine, Chicken/Avian, Hamster, Porcine, Xenopus
- Host
- Rat
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Supportive validation
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- Experimental details
- Western blot analysis of GRP94 in HeLa cell lysate. Proteins were transferred to a nitrocellulose membrane and blocked with 5% non-fat dry milk in PBST for at least 1 hour. The membrane was probed with GRP94/HSP90B1 antibody [9G10] at a dilution of 1:1000 overnight at 4°C on a rocking platform, washed in TBS-0.1%Tween-20, and probed with an HRP-conjugated secondary antibody. Chemiluminescent detection was performed.
- Validation comment
- WB
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- GeneTex (provider)
- Main image
- Experimental details
- WB analysis of HeLa cell lysates using GRP94 antibody [9G10]. Proteins were transferred to a nitrocellulose membrane and blocked with 5% non-fat dry milk in PBST for at least 1 hour. The protein was detected by GRP94 antibody [9G10] at a dilution of 1:1000.
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- GeneTex (provider)
- Main image
- Experimental details
- Western blot analysis of Glucose-Regulated Protein 94 (Grp94) was performed by loading 2-fold serial dilutions of HeLa cell lysate, starting at 10 ug, per well onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a nitrocellulose membrane and blocked with 5% non-fat dry milk in PBST for at least 1 hour. The membrane was probed with a Grp94 monoclonal antibody antibody (GTX22791) at a dilution of 1:1000 overnight at 4ºC on a rocking platform, washed in TBS-0.1%Tween-20, and probed with an HRP-conjugated goat anti-rat IgG secondary antibody at a dilution of 1:25,000 for at least 1 hour. Chemiluminescent detection was performed using ECL Substrate.
- Validation comment
- WB
Supportive validation
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- GeneTex (provider)
- Main image
- Experimental details
- ICC/IF analysis of C6, HeLa and NIH3T3 cells using GRP94 antibody [9G10] (green), Phalloidin (red) and DAPI (blue).
Supportive validation
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- GeneTex (provider)
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- Experimental details
- IHC-P analysis of mouse liver tissue using GRP94 antibody [9G10]. To expose target proteins, heat induced antigen retrieval was performed using 10 mM sodium citrate (pH6.0) buffer, microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:200 with or without GRP94/HSP90B1 antibody [9G10] overnight at 4¢XC in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP, followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.
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- GeneTex (provider)
- Main image
- Experimental details
- IHC-P analysis of poorly differentiated adenocarcinoma from human colon tissue using GRP94 antibody [9G10]. To expose target proteins, heat-induced epitope retrieval was performed using 10 mM sodium citrate (pH 6.0) buffer for 20 minutes at 95oC. Following antigen retrieval, tissues were blocked in 3% BSA in PBST for 30 minutes at room temperature and then probed with GRP94 antibody [9G10] at a dilution of 1:100 for 1 hour in a humidified chamber (right panel). As a negative control, the primary antibody was eliminated from the staining procedure (left panel). Tissues were washed extensively with PBS/0.025% Tween-20 and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection. Tissues were counterstained with hematoxylin and prepped for mouting. Images were taken on a Zeiss Axiovision microscope at 40X magnification.