WB
Western blot analysis of Hsp90 on different lysates with Rabbit anti-Hsp90 antibody at 1/10000 dilution. Lane 1: HeLa cell lysate (20 µg/Lane), Lane 2: A549 cell lysate (20 µg/Lane), Lane 3: NIH/3T3 cell lysate (20 µg/Lane), Lane 4: C6 cell lysate (20 µg/Lane), Lane 5: COS-1 cell lysate (20 µg/Lane), Lane 6: Mouse testis tissue lysate (40 µg/Lane), Lane 7: Rat testis tissue lysate (40 µg/Lane), Exposure time: 4 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/10000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50000 dilution was used for 1 hour at room temperature.ICC/IF
Immunocytochemistry analysis of NIH/3T3 cells labeling Hsp90 with Rabbit anti-Hsp90 antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Hsp90 antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.ICC/IF
Immunocytochemistry analysis of COS-1 cells labeling Hsp90 with Rabbit anti-Hsp90 antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Hsp90 antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1000 dilution.IHC
Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-Hsp90 antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.IHC
Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-Hsp90 antibody at 1/1000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.IHC
Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-Hsp90 antibody at 1/1000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.FC
Flow cytometric analysis of HeLa cells labeling Hsp90. Cells were fixed and permeabilized. Then stained with the primary antibody (1/1000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).IP
Hsp90 was immunoprecipitated from 0.2 mg HeLa cell lysate with Rabbit anti-Hsp90 antibody at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using Rabbit anti-Hsp90 antibody at 1/1000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5000 dilution was used for 1 hour at room temperature. Lane 1: HeLa cell lysate (input), Lane 2:Rabbit anti-Hsp90 antibody IP in HeLa cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-Hsp90 antibody in HeLa cell lysate, Blocking/Dilution buffer: 5% NFDM/TBST, Exposure time: 8 seconds.| Product Name | Hsp90 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Recombinant protein within human Hsp90 beta aa 501-724 / 724. |
| Clonality | monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIHCIPWB |
| WB:1:0000 ICC/IF:1:100 IHC:1:1000 FC:1:1000 IP:1-2μg/sample |
|
| Species Reactivity | HumanMonkeyMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Gene Symbol | HSP90AA1 |
|---|---|
| Gene Synonyms | EL52 HSPN LAP2 HSP86 HSPC1 HSPCA Hsp89 Hsp90 LAP-2 HSP89A HSP90A HSP90N Hsp103 HSPCAL1 HSPCAL4 HEL-S-65p |
| Gene Full Name | heat shock protein 90 alpha family class A member 1 |
| Gene Summary | The protein encoded by this gene is an inducible molecular chaperone that functions as a homodimer. The encoded protein aids in the proper folding of specific target proteins by use of an ATPase activity that is modulated by co-chaperones. Two transcript variants encoding different isoforms have been found for this gene. [provided by RefSeq, Jan 2012] |
| Alternative Names | Heat shock 86 kDa antibody Heat shock protein 90kDa alpha cytosolic class A member 1 antibody Heat shock protein 90kDa alpha cytosolic class B member 1 antibody Heat shock protein HSP 90 alpha antibody Heat shock protein HSP 90 beta antibody Heat shock protein HSP 90-alpha antibody HS90A_HUMAN antibody HSP 84 antibody HSP 86 antibody Hsp 90 antibody Heat shock 86 kDa antibody Heat shock protein 90kDa alpha cytosolic class A member 1 antibody Heat shock protein 90kDa alpha cytosolic class B member 1 antibody Heat shock protein HSP 90 alpha antibody Heat shock protein HSP 90 beta antibody Heat shock protein HSP 90-alpha antibody HS90A_HUMAN antibody HSP 84 antibody HSP 86 antibody Hsp 90 antibody HSP86 antibody HSP90A antibody HSP90AA1 antibody HSP90AB1 antibody HSP90B antibody HSPC1 antibody HSPC2 antibody HSPCAL1 antibody HSPCAL4 antibody Renal carcinoma antigen NY-REN-38 antibody |
| Molecular Weight(MW) | 85kDa(Observed band size: 85kDa) |
| Cellular Localization | Nucleus, Cytoplasm, Melanosome, Cell membrane, Mitochondrion. |
WB
Western blot analysis of Hsp90 on different lysates with Rabbit anti-Hsp90 antibody at 1/10000 dilution. Lane 1: HeLa cell lysate (20 µg/Lane), Lane 2: A549 cell lysate (20 µg/Lane), Lane 3: NIH/3T3 cell lysate (20 µg/Lane), Lane 4: C6 cell lysate (20 µg/Lane), Lane 5: COS-1 cell lysate (20 µg/Lane), Lane 6: Mouse testis tissue lysate (40 µg/Lane), Lane 7: Rat testis tissue lysate (40 µg/Lane), Exposure time: 4 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody at 1/10000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50000 dilution was used for 1 hour at room temperature.
ICC/IF
Immunocytochemistry analysis of NIH/3T3 cells labeling Hsp90 with Rabbit anti-Hsp90 antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Hsp90 antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.
ICC/IF
Immunocytochemistry analysis of COS-1 cells labeling Hsp90 with Rabbit anti-Hsp90 antibody at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Hsp90 antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1000 dilution.
IHC
Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-Hsp90 antibody at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
IHC
Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-Hsp90 antibody at 1/1000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
IHC
Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-Hsp90 antibody at 1/1000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
FC
Flow cytometric analysis of HeLa cells labeling Hsp90. Cells were fixed and permeabilized. Then stained with the primary antibody (1/1000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
IP
Hsp90 was immunoprecipitated from 0.2 mg HeLa cell lysate with Rabbit anti-Hsp90 antibody at 2 µg/25 µl agarose. Western blot was performed from the immunoprecipitate using Rabbit anti-Hsp90 antibody at 1/1000 dilution. Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5000 dilution was used for 1 hour at room temperature. Lane 1: HeLa cell lysate (input), Lane 2:Rabbit anti-Hsp90 antibody IP in HeLa cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-Hsp90 antibody in HeLa cell lysate, Blocking/Dilution buffer: 5% NFDM/TBST, Exposure time: 8 seconds.| Application Notes | WB:1:0000 ICC/IF:1:100 IHC:1:1000 FC:1:1000 IP:1-2μg/sample |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term. |
| Storage Buffer | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
