GAD65/GAD2 Antibody (monoclonal, 4E12)

• Catalog Number: orb654393
• Category: Antibodies
• Description: GAD65/GAD2 Antibody (monoclonal, 4E12)
• Species/Host: Mouse
• Clonality: Monoclonal
• Clone Number: 4E12
• Tested applications: WB
• Reactivity: Mouse, Rat
• Isotype: Mouse IgG1
• Immunogen: A synthetic peptide corresponding to a sequence at the N-terminus of human GAD65 (131-164aa KVIDFHYPNELLQEYNWELADQPQNLEEILMHCQ), different from the related mouse and rat sequences by one amino acid.
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Dilution range: Western blot, 0.1-0.5μg/ml, Mouse, Rat
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 65 kDa
• UniProt ID: Q05329
• Storage: Store 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 freeze-thaw cycles.
• 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. Add 0.2ml of distilled water will yield a concentration of 500ug/ml.
• Expiration Date: 12 months from date of receipt.

Western blot analysis of GAD65/GAD2 using anti-GAD65/GAD2 antibody (orb654393). 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 50ug of sample under reducing conditions. Lane 1: rat brain tissue lysates, Lane 2: mouse brain tissue lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA 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-GAD65/GAD2 antigen affinity purified monoclonal antibody (Catalog # orb654393) 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 GAD65/GAD2 at approximately 65KD. The expected band size for GAD65/GAD2 is at 65KD.