Antibody was affinity purified using an epitope specific to CBX8 immobilized on solid support. The epitope recognized by A300-882A maps to a region between residue 350 and the C-terminus (residue 389) of human Chromobox homolog 8 using the numbering given in entry NP_065700.1 (GeneID 57332). Immunoglobulin concentration was determined by extinction coefficient: absorbance at 280 nm of 1.4 equals 1.0 mg of IgG.
chromobox homolog 8 (Pc class homolog, Drosophila), chromobox protein homolog 8, hPc3, Pc class 3 homolog, PC3, polycomb 3, Polycomb 3 homolog, RC1, rectachrome 1
between 350 and C-term
2 - 8°C
Detection of human CBX8 by western blot. Samples: Whole cell lysate (50 µg) from HeLa and HEK293T cells prepared using NETN lysis buffer. Antibody: Affinity purified rabbit anti-CBX8 antibody A300-882A (lot A300-882A-2) used for WB at 0.1 µg/ml. Detection: Chemiluminescence with an exposure time of 30 seconds.
Detection of human CBX8 by western blot of immunoprecipitates. Samples: Whole cell lysate (0.5 or 1.0 mg per IP reaction; 20% of IP loaded) from HeLa cells prepared using NETN lysis buffer. Antibodies: Affinity purified rabbit anti-CBX8 antibody A300-882A (lot A300-882A-2) used for IP at 6 µg per reaction. CBX8 was also immunoprecipitated by rabbit anti-CBX8 antibody BL3751. For blotting immunoprecipitated CBX8, A300-882A was used at 0.4 µg/ml. Detection: Chemiluminescence with an exposure time of 30 seconds.
Localization of CBX8 Binding Sites by ChIP-sequencing. Chromatin from K562 cells was immunoprecipitated with anti-CBX8 antibody A300-882A and analyzed by DNA sequencing. The figure illustrates the peak distribution of CBX8 binding within a 4 Mb region of chromosome 1 as detected using anti-CBX8 antibody A300-882A. ChIP-seq validation performed by Diogenode, Denville, NJ.