SP-A affinity purified antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to a sequence at the C-terminal of Human SP-A.
Anti-SP-A is tested for Western Blotting. Specific conditions for reactivity should be optimized by the end user. Expect a band approximately ~26.2 kDa corresponding to the appropriate cell lysate or extract.
Western blot analysis of SP-A/SFTPA1 using anti-SFTPA1 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 50ug of sample under reducing conditions. Lane 1: rat lung 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 affinity purified rabbit anti-SFTPA1 antigen polyclonal antibody at 0.5 µg/mL overnight at 4C, 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 1:10000 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 SFTPA1 at approximately 26-35KD. The expected band size for SFTPA1 is at 26KD.
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