Anti-Hsp90 beta/HSP90AB1 Antibody (monoclonal, 7B7F5)

• Catalog Number: orb1474875
• Category: Antibodies
• Description: Anti-Hsp90 beta/HSP90AB1 Antibody (monoclonal, 7B7F5). Tested in Flow Cytometry, IF, ICC, WB applications. This antibody reacts with Human, Mouse, Rat.
• Species/Host: Mouse
• Clonality: Monoclonal
• Clone Number: 7B7F5
• Tested applications: FC, ICC, IF, WB
• Reactivity: Human, Mouse, Rat
• Isotype: IgG2b
• Immunogen: A synthetic peptide corresponding to a sequence at the C-terminus of human Hsp90 beta, identical to the related mouse and rat sequences.
• Antibody Type: Primary Antibody
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 90 kDa
• UniProt ID: P08238
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Alternative names: A32 antigen antibody; CD 146 antibody; CD146 antib
• Note: For research use only
• Application notes: Western blot, 0.25-0.5 μg/ml, Human, Mouse, Rat Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human Flow Cytometry (Fixed), 1-3 μg/1x106 cells, Human. Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml
• Expiration Date: 12 months from date of receipt.

Flow Cytometry analysis of CACO-2 cells using anti-Hsp90 beta/HSP90AB1 antibody. Overlay histogram showing CACO-2 cells (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 mouse anti-Hsp90 beta/HSP90AB1 Antibody (1 µg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (5-10 µg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1 µg/1x10^6) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

IF analysis of Hsp90 beta/HSP90AB1 using anti-Hsp90 beta/HSP90AB1 antibody. Hsp90 beta/HSP90AB1 was detected in an immunocytochemical section of MCF-7 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 µg/mL mouse anti-Hsp90 beta/HSP90AB1 Antibody overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG 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.

Western blot analysis of Hsp90 beta/HSP90AB1 using anti-Hsp90 beta/HSP90AB1 antibody. 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 Raji whole cell lysates, Lane 4: human A431 whole cell lysates, Lane 5: rat brain tissue lysates, Lane 6: rat heart tissue lysates, Lane 7: mouse brain tissue lysates, Lane 8: 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 mouse anti-Hsp90 beta/HSP90AB1 antigen affinity purified monoclonal antibody 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-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for Hsp90 beta/HSP90AB1 at approximately 90 kDa. The expected band size for Hsp90 beta/HSP90AB1 is at 84 kDa.