Anti-DRIP-130 detects a 130 kDa band by immunoblot analysis using a dilution of 1:1,000. For immunoprecipitation a dilution of 1:100 or 1:200 is suggested; pre-clearing with a non-specific rabbit IgG is helpful to reduce background. Optimal titers for applications should be determined by the researcher. anti-DRIP130 has been used to detect DRIP130 using nuclear extracts from mouse thymus, spleen, and brain. Unfortunately, we have not tested reactivity on any cell line extracts. The following standard procedure is recommended: separate 10 micrograms of thymus nuclear extract by 7.5% SDS-PAGE (0.75 to 1 mm thick). Transfer to nitrocellulose without SDS in the transfer buffer (pre-equilibrate the gel in transfer buffer for 30 min prior to transfer). Blocking is performed with 5% non-fat dry milk in TTBS for 1 hr at room temperature. Incubate anti-DRIP130 antibody at a 1:1,000 for 1 hr at room temperature. Dilute Donkey anti-Rabbit IgG-HRP 1:5,000 and react 1 hr at room temperature. A predominant band at ~130 KDa is detected under these conditions. Control or normal rabbit serum fails to recognize the 130 kDa protein.