CD14 affinity purified antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to an internal region of mouse CD14.
Anti-CD14 is tested for Western Blotting. Specific conditions for reactivity should be optimized by the end user. Expect a band approximately ~39.2 kDa corresponding to the appropriate cell lysate or extract.
Western blot analysis of CD14 using anti-CD14 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 30µg of sample under reducing conditions. Lane 1: mouse RAW264.7 whole cell 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 affinity purified rabbit anti-CD14 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 1:5000 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 CD14 at approximately 50 kDa. The expected band size for CD14 is at 39 kDa.
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