Antibody was affinity purified using an epitope specific to hSET1 immobilized on solid support. The epitope recognized by A300-289A maps to a region between residues 1200 and 1250 of human hSET1 (KIAA0339) using the numbering given in TrEMBL entry O15047 (GeneID 9739). Immunoglobulin concentration was determined by extinction coefficient: absorbance at 280 nm of 1.4 equals 1.0 mg of IgG.
histone-lysine N-methyltransferase SETD1A, hSET1A, KMT2F, lysine N-methyltransferase 2F, SET domain-containing protein 1A, Set1, set1/Ash2 histone methyltransferase complex subunit SET1, Set1A
between 1200 and 1250
2 - 8°C
Detection of human hSET1 by western blot of immunoprecipitates. Samples: Nuclear Extract (1 mg for IP; 20% of IP loaded) from HEK293T cells. Antibodies: Affinity purified rabbit anti-hSET1 antibody A300-289A (lot A300-289A-4) used for IP at 6 µg per reaction. hSET1 was also immunoprecipitated by rabbit anti-hSET1 antibody BL1192. For blotting immunoprecipitated hSET1, A300-289A was used at 0.4 µg/ml. Detection: Chemiluminescence with an exposure time of 3 minutes.
ChIP-chip scatter plot of anti-hSET1 (A300-289A) enriched DNA binding sites versus input reference DNA. A. 10 µg of A300-289A was used to immunoprecipitate chromatin from K-562 cells according to Ren et al (Genes Dev. 2002 16: 245-256). immunoprecipitatesd DNA and reference DNA were amplified via ligation-mediated PCR and the products labeled with fluorescent dUTPs. The labeled ChIP and reference DNA were pooled, hybridized to a DNA microarray, and analyzed. Data points below the +3 SD curve (red line) represent significantly enriched binding sites. B. As a control, a similar experiment was performed using normal rabbit IgG. Compared to the anti-hSET1 ChIP, normal rabbit IgG showed little enrichment.