Anti-Hsp90 alpha/HSP90AA1 Antibody Picoband®

IHC analysis of Hsp90 Alpha using anti-Hsp90 Alpha antibody (PB9089).
Hsp90 Alpha was detected in a paraffin-embedded section of human intestinal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-Hsp90 Alpha Antibody (PB9089) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # SA1022) with DAB as the chromogen.

IHC analysis of Hsp90 Alpha using anti-Hsp90 Alpha antibody (PB9089).
Hsp90 Alpha was detected in a paraffin-embedded section of mouse brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-Hsp90 Alpha Antibody (PB9089) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

ICC analysis of Hsp90 Alpha using anti-Hsp90 Alpha antibody (PB9089).
Hsp90 Alpha was detected in an immunocytochemical section of A549 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 1 μg/ml rabbit anti-Hsp90 Alpha Antibody (PB9089) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

IHC analysis of Hsp90 Alpha using anti-Hsp90 Alpha antibody (PB9089).
Hsp90 Alpha was detected in a paraffin-embedded section of rat brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-Hsp90 Alpha Antibody (PB9089) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

ICC analysis of Hsp90 Alpha using anti-Hsp90 Alpha antibody (PB9089).
Hsp90 Alpha was detected in an immunocytochemical section of MCF-7 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 1 μg/ml rabbit anti-Hsp90 Alpha Antibody (PB9089) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

Western blot analysis of Hsp90 Alpha using anti-Hsp90 Alpha antibody (PB9089).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human Jurkat whole cell lysates,
Lane 3: human A431 whole cell lysates,
Lane 4: rat brain tissue lysates,
Lane 5: rat heart tissue lysates,
Lane 6: mouse brain tissue lysates,
Lane 7: mouse heart tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Hsp90 Alpha antigen affinity purified polyclonal antibody (Catalog # PB9089) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Hsp90 Alpha at approximately 95 kDa. The expected band size for Hsp90 Alpha is at 85 kDa.

IF analysis of Hsp90 alpha using anti-Hsp90 alpha antibody (PB9089).
Hsp90 alpha was detected in immunocytochemical section of MCF-7 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5μg/mL rabbit anti-Hsp90 alpha Antibody (PB9089) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Flow Cytometry analysis of A549 cells using anti-Hsp90 alpha antibody (PB9089).
Overlay histogram showing A549 cells stained with PB9089 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Hsp90 alpha Antibody (PB9089, 1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Effect of BC on the expression of diverse proteins in BT549 cells. (A) Image of MAPK signaling pathway-related protein expression bands. (B) The expression of MAPK signaling pathway-related proteins after BC treatment of BT549 cells. (* P < 0.05, ** P < 0.01 versus control group). (C) Molecular docking diagram of BC and HSP90AA1 protein. (D) Molecular docking diagram of BC and PTGS21 protein. (E) Image of hub protein expression bands. (F) The expression of hub proteins after BC treatment of BT549 cells. (G) Images of HSP90AA1, PTGS2, and β-Actin protein expression bands of the cellular thermal shift assay. (H) Statistical results of HSP90AA1, PTGS2, and β-Actin protein expression bands of the cellular thermal shift assay. (* P < 0.05, ** P < 0.01 versus control group).
Index in PubMed under a CC BY license. PMID: 39502536

HSP90AA1 and PTGS2 gene mutation and methylation in BRCA. (A, B) The alteration frequency, mutation types, mutation sites, and promoter methylation level of HSP90AA1 and PTGS2. (C) The relationship between SNV, CNV and HSP90AA1 expression, and the correlation between HSP90AA1 and the expression of 44 marker genes of RNA modification. (D) The relationship between SNV, CNV and PTGS2 expression, and the correlation between PTGS2 and the expression of 44 marker genes of RNA modification. (* P < 0.05, **** P < 0.0001 versus control group).
Index in PubMed under a CC BY license. PMID: 39502536

Expression of hub genes in BCRA from TCGA and GTEx data. (A) HSP90AA1 was upregulated in BRCA. (B) The expression of HSP90AA1 was associated with the stage of BRCA. (C) The protein expression of HSP90AA1 in BRCA. (D) PTGS2 was downregulated in BRCA. (E) The expression of PTGS2 was associated with the stage of BRCA. (F) The protein expression of PTGS2. (* P < 0.05 and **** P < 0.0001 versus control group).
Index in PubMed under a CC BY license. PMID: 39502536