Alpha A Crystallin Antibody (monoclonal, 10B9)

• Catalog Number: orb763150
• Category: Antibodies
• Description: Alpha A Crystallin Antibody (monoclonal, 10B9)
• Species/Host: Mouse
• Clonality: Monoclonal
• Clone Number: 10B9
• Tested applications: ICC, IF, WB
• Reactivity: Human, Mouse, Rat
• Isotype: Mouse IgG1
• Immunogen: E. coli-derived human Alpha A Crystallin recombinant protein (Position: M1-S173). Human Alpha A Crystallin shares 94.8% amino acid (aa) sequence identity with both mouse and rat Alpha A Crystallin.
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Dilution range: Western blot, 0.25-0.5 μg/ml, Mouse, Rat Immunocytochemistry/Immunofluorescence, 5 μg/ml, Human
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 20-23 kDa
• UniProt ID: P02489
• Storage: At -20°C for one year from date of receipt. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for six months. Avoid repeated freezing and thawing.
• Note: For research use only
• Application notes: Tested Species: In-house tested species with positive results. Other applications have not been tested. Optimal dilutions should be determined by end users. Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Expiration Date: 12 months from date of receipt.

Western blot analysis of Alpha A Crystallin using anti-Alpha A Crystallin antibody (orb763150). 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: rat eye tissue lysates, Lane 2: mouse eye 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-Alpha A Crystallin antigen affinity purified monoclonal antibody (Catalog # orb763150) 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 (Catalog # orb90502) with Tanon 5200 system. A specific band was detected for Alpha A Crystallin at approximately 20-23 kDa. The expected band size for Alpha A Crystallin is at 20-23 kDa.

IF analysis of Alpha A Crystallin using anti-Alpha A Crystallin antibody (orb763150). Alpha A Crystallin was detected in an immunocytochemical section of HepG2 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (orb90553) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 5 μg/mL mouse anti-Alpha A Crystallin Antibody (orb763150) 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.