This affinity purified antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to an internal sequence of human Cripto-1 protein.
This affinity purified antibody has been tested for use in ELISA and western blotting. Specific conditions for reactivity should be optimized by the end user. Expect a band approximately 21 kDa in size corresponding to Cripto-1 by western blotting in the
Western blot using Rocklands affinity purified anti-Cripto-1 antibody shows detection of endogenous and transfected Cripto-1 from mouse and human sources. The Cripto-1 band appears above the 17.8 kDa marker. Endogenous detection is shown using mouse P19 embryonal carcinoma cells and human NCCIT testicular embryonal carcinoma cells. EpH4 CR-1 is a mouse mammary epithelial cell line stably expressing mouse Cripto-1. NTERA cells are human embryonal carcinoma cells that, when overgrown, differentiate and lose Cripto-1 expression. COS7 cells transfected with human Cripto-1 expression vector were used as a positive control and EpH4 WT cells were used as a negative control. A non-specific band at 45kDa may be present in some preparations. The primary antibody was used at a 1:500 dilution. Personal communication, C. Bianco, NCI, Bethesda, MD
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