CHIPseq abreview for Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade

Excellent
Abreviews
Application
CHIPseq
Sample
Mouse Cell lysate - nuclear (MEF)
Specification
MEF

Other product details

Incubation time
5 hour(s) and 0 minute(s) · Diluent: 1% BSA in PBS
Dilution
1/1000

Secondary antibody

Secondary antibody
None used

Additional data

Additional Notes
The real application is sequential ChIP or re-ChIP. First IP was perfomed with anti H3 and later on Str Beads were used to do a new IP, before QT-PCR analysis. Figure legend: H3 oxidation in major satellite region was determined by sequential ChIP (re-ChIP) in control and SNA1 KO MEFs. The lysate was incubated with biotin-hydrazide (an activated biotin that reacts with oxidized H3) before re-ChIP. Extracts were sequentially immunoprecipitated with anti-H3 and streptavidin beads. DNA binding was quantified by qPCR. Data were normalized to the total amount of H3 immunoprecipitated and to the input and expressed as fold enrichment over the data obtained when an irrelevant IgG was used. Brief Protocol: To detect H3 oxidation, ChIP assays were performed as described previously (Herranz et al., 2012). Briefly, cell extracts were incubated with activated biotin, crosslinked, and immunoprecipitated with anti-histone H3 immunocomplexes were reextracted with SDS lysis buffer and reimmunoprecipitated with streptavidinmagnetic beads for 30 min at 4C. Samples were then treated with elution buffer and incubated at 65C to reverse formaldehyde crosslinking. Results were quantified by taking into account the total amount of H3 immunoprecipitated in each condition.
PubMed ID
24239292: View

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Submitted Jan 30 2014

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