MA3-011

antibody from Invitrogen Antibodies

Targeting: HSP90AA1 FLJ31884, Hsp89, Hsp90, HSP90N, HSPC1, HSPCA

Provider product page for MA3-011

Western blot

Immunocytochemistry

Immunoprecipitation

Immunohistochemistry

Other assay

Antibody data

Antibody Data
Antigen structure
References [64]
Comments [0]
Validations
Western blot [2]
Immunocytochemistry [4]
Immunohistochemistry [3]
Other assay [2]

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Product number: MA3-011 - Provider product page
Provider: Invitrogen Antibodies
Product name: HSP90 alpha Monoclonal Antibody (3G3)
Antibody type: Monoclonal
Antigen: Purifed from natural sources
Description: MA3-011 detects heat shock protein 90 kDa (HSP90) from human, mouse, rat, rainbow trout, and chicken tissues. MA3-011 has been successfully used in Western blot, immunofluorescence, immunohistochemistry, and immunoprecipitation procedures. By Western blot, this antibody detects a 90 kDa protein representing HSP90 from Hepa 1 cell extract. It has been reported that up to 90% of cytosolic HSP90 can be immunoprecipitated with this product under low ionic strength conditions. For Western blots using MA3-011, nitrocellulose is recommended. PVDF membranes typically do not yield acceptable results with this antibody. Before use, centrifuge suspension (4000 g for 15 minutes) and discard supernatant. Resuspend pellet in 200 µL of PBS, pH 7.2. If desired, 0.05% sodium azide may be added to resuspended solution. Removal of residual ammonium sulfate is not usually necessary for immunoprecipitation and Western blot procedures, however, any other uses may require its removal by dialysis or equivalent method. Stable for 6 months after PBS resuspension. The MA3-011 antigen is purified HSP90 from Hepa 1 cytosol.
Reactivity: Human, Mouse, Rat, Chicken/Avian
Host: Mouse
Isotype: IgM
Antibody clone number: 3G3
Vial size: 400 µL
Concentration: Conc. Not Determined
Storage: -20° C, Avoid Freeze/Thaw Cycles

HSP90AA1 protein structure - MA3-011 shown in red.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Heat Shock Protein 90 using Heat Shock Protein 90 Monoclonal antibody (3G3) (Product # MA3-011) shows staining in C6 glioma cells. Heat Shock Protein 90 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing Heat Shock Protein 90 (Product # MA3-011) at a dilution of 1:200 over night at 4 C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody (Product # 35552 for GAR, Product # 35503 for GAM). Images were taken at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Heat Shock Protein 90 using Heat Shock Protein 90 Monoclonal antibody (3G3) (Product # MA3-011) shows staining in HeLa cells. Heat Shock Protein 90 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing Heat Shock Protein 90 (Product # MA3-011) at a dilution of 1:200 over night at 4 C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody (Product # 35552 for GAR, Product # 35503 for GAM). Images were taken at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Heat Shock Protein 90 using Heat Shock Protein 90 Monoclonal antibody (3G3) (Product # MA3-011) shows staining in WiDr colon carcinoma cells. Heat Shock Protein 90 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with or an antibody recognizing Heat Shock Protein 90 (Product # MA3-011) at a dilution of 1:200 over night at 4 C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody (Product # 35552 for GAR, Product # 35503 for GAM). Images were taken at 60X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of Heat Shock Protein 90 (Hsp90) (green) in HeLa cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA (Product # 37525) for 15 minutes at room temperature. Cells were probed without (left panel) or with (right panel) a Hsp90 monoclonal antibody (Product # MA3-011), at a dilution of 1:100 for at least 1 hour at room temperature, washed with PBS, and incubated with a fluorescently-conjugated goat anti-mouse IgM secondary antibody at a dilution of 1:400 for 30 minutes at room temperature. F-Actin (red) was stained with DyLight-554 Phalloidin (Product # 21834) and nuclei (blue) were stained with Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific ArrayScan at 20X magnification.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry was performed on cancer biopsies of deparaffinized Human breast carcinoma tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM 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 a mouse monoclonal antibody recognizing Heat Shock Protein 90 (Product # MA3-011) or without primary antibody (negative control) overnight at 4°C 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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry was performed on normal biopsies of deparaffinized Human placenta tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM 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 a mouse monoclonal antibody recognizing Heat Shock Protein 90 (Product # MA3-011) or without primary antibody (negative control) overnight at 4°C 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.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunohistochemistry was performed on normal biopsies of deparaffinized Human tonsil tissue. To expose target proteins, heat induced antigen retrieval was performed using 10mM 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 a mouse monoclonal antibody recognizing Heat Shock Protein 90 (Product # MA3-011) or without primary antibody (negative control) overnight at 4°C 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.