Alternate Name: CHAT, CMS1A, CMS1A2, choline acetyltransferase, acetyl CoA:choline O-acetyltransferase Domain Information: Gene Ontology Terms: Application(s): Peptide ELISA: antibody detection limit dilution 1:32,000. Western Blot: Approx 70kD band observed in Human Placenta lysates (calculated MW of 70.5kD according to NP_065574 and NP_066264). Recommended concentration: 0.03-0.3ug/ml. 0.03ug/ml staining of Human Placenta lysate (35ug protein in RIPA buffer). Primary incubation was 1 hour. Detected by chemiluminescence. Optimal dilutions to be determined by the researcher. Storage and Stability: May be stored at 4C for short-term only. For long-term storage and to avoid repeated freezing and thawing, aliquot and store at -20C. Aliquots are stable for at least 12 months at -20C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer. Peptide Blocking: Corresponding peptide is available for Peptide Blocking studies. See C5057-06A-P. Antibody is typically 0.5mg/ml and peptide is supplied as a 100ul pellet. When peptide is reconstituted in 200ul water, the concentration would also be 0.5mg/ml. To start, the best ratio would be 1:1 (molar excess of peptide relative to antibody when identical volumes are mixed). Mix equal volumes of peptide and antibody at the required dilution and leave at ambient temperature. It is best is to have two identical blots to be incubated with equal amount of antibody, but one with the antibody pre-adsorbed to the peptide for 20min. Then incubate and develop the two blots in parallel.
