The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use 2.5µg for 107 cells.
Use a concentration of 1 µg/ml. Detects a band of approximately 109 kDa (predicted molecular weight: 109 kDa).
RelevanceChp1 a chromodomain protein associates with the S. pombe mating type locus and telomeres. Chp1 localization to heterochromatin depends on its chromodomain and the C-terminal domain of the protein. Chp1 and centromeric siRNAs are components of the RITS (RNA-induced initiation of transcriptional gene silencing) complex. The association of RITS with centromeres is linked to Dicer activity, which is required to cleave centromeric transcripts to produce short interfering RNAs that actively recruit components of heterochromatin to centromeres.
Western blot - Anti-Chp1 antibody - ChIP Grade (ab18191)This image is courtesy of Danesh Moazed's Laboratory, Harvard, Boston.
All lanes : Anti-Chp1 antibody - ChIP Grade (ab18191) at 1 µg/ml
Lane 1 : S. pombe chp1Δ lysate Lane 2 : S. pombe wildtype lysate
Predicted band size : 109 kDa Observed band size : 109 kDa Additional bands at : 102 kDa (possible degradation product).
This image is courtesy of Danesh Moazed's Laboratory, Harvard, Boston.
ab18191 specifically recognises Chp1 at 109 kDa in wildtype S. pombe lysate but not in chp1Δ lysate.
ChIP - Chp1 antibody - ChIP Grade (ab18191)
X-ChIP performed with ab18191 on Schizosaccharomyces pombe Cell lysate (whole cell). 5µl antibody were used per 500µl of chromatin/cell lysate according to standard S. pombe ChIP protocols. Protein-A Sepharose was used to recover the ChIPs. In the positive control the researcher specifically induced silencing of the ura4+ gene. For the negative control the researcher performed multiplex PCR with primers amplifying ura4+ and an un-silenced gene. In addition, act1+ was amplified in a separate PCR reaction. Chp1 only binds to ura4+ (and only if silencing is induced).
This image is courtesy of an Abreview submitted by Marc B? on 9 February 2006