Anti-Hsp90 beta 抗体

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Hsp90 beta antibody (AB2927)

Immunocytochemistry/Immunofluorescence analysis of HSP90 beta shows staining in U251 cells. HSP 90 beta 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 ab2927 at a dilution of 1 : 100 over night at 4°C, washed with PBS and incubated with a DyLight-488 conjugated goat anti-rabbit secondary antibody. Images were taken at 60X magnification.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp90 beta antibody (AB2927)

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 : 100 with a rabbit polyclonal antibody recognizing Heat Shock Protein 84 ab2927 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.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp90 beta antibody (AB2927)

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 : 20 with a rabbit polyclonal antibody recognizing Heat Shock Protein 84 ab2927 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.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Hsp90 beta antibody (AB2927)

Immunocytochemistry/Immunofluorescence analysis of HSP90 beta shows staining in HepG2 cells. HSP 90 beta 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 ab2927 at a dilution of 1 : 100 over night at 4°C, washed with PBS and incubated with a DyLight-488 conjugated goat anti-rabbit secondary antibody. Images were taken at 60X magnification.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Hsp90 beta antibody (AB2927)

Immunocytochemistry/Immunofluorescence analysis of HSP90 beta (green) in HeLa and NIH3T3 cells. Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature. Cells were blocked with 1% BSA for 15 minutes at room temperature. Cells were incubated with ab2927 at a dilution of 1 : 50 for at least 1 hour at room temperature, washed with PBS, and incubated with a DyLight 488 goat-anti-rabbit IgG secondary antibody (1 : 400) for 30 minutes at room temperature. Nuclei (blue) were stained with Hoechst 33342 dye. Images were taken at 20X magnification.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp90 beta antibody (AB2927)

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 : 100 with a rabbit polyclonal antibody recognizing Heat Shock Protein 84 ab2927 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.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Hsp90 beta antibody (AB2927)

Immunocytochemistry/Immunofluorescence analysis of HSP90 beta shows staining in A2058 cells. HSP 90 beta 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 ab2927 at a dilution of 1 : 100 over night at 4°C, washed with PBS and incubated with a DyLight-488 conjugated goat anti-rabbit secondary antibody. Images were taken at 60X magnification.

• Flow Cyt

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Flow Cytometry - Anti-Hsp90 beta antibody (AB2927)

Flow cytometry analysis of HSP90 was done on HeLa cells. Cells were fixed, permeabilized and stained with a HSP90 rabbit polyclonal antibody (ab2927) (blue histogram) or a rabbit IgG isotype control (black histogram) at a dilution of 10 μg/mL. After incubation for 1 hour on ice, the cells were labeled with a Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor^® 647 conjugate at a dilution of 1/50 for 1 hour on ice. A representative 10,000 cells were acquired and analyzed for each sample.

• IP

Unknown

Immunoprecipitation - Anti-Hsp90 beta antibody (AB2927)

Immunoprecipitation of Hsp90 was performed on HeLa cells. Antigen : antibody complexes were formed by incubating 500μg whole cell lysate with 2μg of Hsp90 polyclonal antibody (ab2927) overnight on a rocking platform at 4°C. Immune complexes were captured on 50μl Protein A/G Agarose washed extensively and eluted with Buffer. Samples were resolved on a 4-20% Tris-HCl polyacrylamide gel, transferred to a PVDF membraneand blocked with 5% BSA/TBST for at least 1 hour. The membrane was probed with a Hsp90 polyclonal antibody (ab2927) at a dilution of 1 : 1000 overnight rotating at 4°C, washed in TBSTand probed with HRP detection reagent at a dilution of 1 : 1000 for at least one hour. Chemiluminescent detection was performed.

All lanes:

Immunoprecipitation - Anti-Hsp90 beta antibody (ab2927)

Predicted band size: 83 kDa

false

• WB

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Western blot - Anti-Hsp90 beta antibody (AB2927)

Samples were loaded onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with 5% BSA/TBST for at least 1 hour. The membrane was then probed with primary antibody (ab2927) overnight at 4°C on a rocking platform, washed in TBS-0.1%Tween 20, and probed with secondary antibody for at least one hour. Chemiluminescent detection was performed using SuperSignal West Pico.

All lanes:

Western blot - Anti-Hsp90 beta antibody (ab2927) at 1/1000 dilution

Lane 1:

MCF7 whole cell lysate at 50 µg

Lane 2:

293T whole cell lysate at 50 µg

Lane 3:

HeLa whole cell lysate at 50 µg

Lane 4:

K562 whole cell lysate at 50 µg

Lane 5:

A431 whole cell lysate at 50 µg

Lane 6:

HepG2 whole cell lysate at 50 µg

Lane 7:

COS7 whole cell lysate at 50 µg

Lane 8:

NIH3T3 whole cell lysate at 50 µg

Lane 9:

NRK whole cell lysate at 50 µg

Secondary

All lanes:

Goat anti-rabbit IgG-HRP at 1/20000 dilution

Predicted band size: 83 kDa

false

• WB

Unknown

Western blot - Anti-Hsp90 beta antibody (AB2927)

Western blot of mouse HSP 86 using ab2927.

All lanes:

Western blot - Anti-Hsp90 beta antibody (ab2927)

Predicted band size: 83 kDa

false

• WB

Supplier Data

Western blot - Anti-Hsp90 beta antibody (AB2927)

All lanes:

Western blot - Anti-Hsp90 beta antibody (ab2927)

Lane 1:

CRISPR targeted HSP90 beta knockout HeLa whole cell lysate

Lane 2:

Wild-type HeLa whole cell lysate

Predicted band size: 83 kDa

false