Anti-MELK (MELK, maternal embryonic leucine zipper kinase, KIAA0175, RP11-8N6.1, HPK38, OTTHUMP00000

Alternate Name: MELK, maternal embryonic leucine zipper kinase, KIAA0175, RP11-8N6.1, HPK38, OTTHUMP00000046113, pEg3 kinase, Domain Information: Kinase associated domain 1, Serine/Threonine protein kinases, catalytic domain , Gene Ontology Terms: protein tyrosine kinase activity, ATP binding, protein serine/threonine kinase activity, protein amino acid phosphorylation, transferase activity, Application(s): Peptide ELISA: (antibody detection limit dilution 1:32,000). Western blot: Approx 120-150kD band seen in Jurkat and HeLa cell lysates. Recommended for use at 0.5-2 ug/ml. Please note that the band we consistently detect in western blots differs from the predicted size of 72kD. Complete blocking by the immunizing peptide is observed with peptide neutralization experiments. It is to be noted that MELK is a member of AMPK family, which has several consensus glycosylation sites. Glycosylations are known to affect migration of proteins on a SDS-PAGE [Hames, B.D and Rickwood, D., eds. Gel electrophoresis of proteins: a practical approach. Oxford University Press]. Further the activity of this antibody was confirmed against recombinant, GST fused MELK protein by Western blotting. 0.5ug/ml staining of Jurkat lysate (RIPA buffer, 1.4E5 cells per lane). Detected by western blot using chemiluminescence. Optimal dilutions to be determined by the researcher. Storage and Stability: May be stored at 4°C for short-term only. For long-term storage and to avoid repeated freezing and thawing, aliquot and store at -20°C. Aliquots are stable for at least 12 months at -20°C. 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 M2867-65A-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.