Anti-Ctip1/BCL-11A antibody [14B5] (ab19487)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
-Add protease inhibitors to all lysis solutions before use.	-Keep cells on ice between the rounds of homogenisations. - Increase or decrease the homogenization step to maximize the yield of nuclei depending on cell line. -Do not place ethidium bromide in the agarose gel or the electrophoresis buffer, because of the presence of SDS.

Upstream tips
-Add protease inhibitors to all lysis solutions before use.
Protocol tips
-Keep cells on ice between the rounds of homogenisations. - Increase or decrease the homogenization step to maximize the yield of nuclei depending on cell line. -Do not place ethidium bromide in the agarose gel or the electrophoresis buffer, because of the presence of SDS.

Publication protocol

The chromatin immunoprecipitation (ChIP) assay followed the general protocol (30). The cell extracts from cross-linked primary human adult erythroid cells were immunoprecipitated with the addition of purified rabbit IgG and antibodies against BCL11A, H3K4me2, H3K9me2 and H3K9me3, respectively. The immunocomplexes were subjected to a reversal of cross-links at 67°C for 4 h and treated with proteinase K at 45°C overnight. The DNA samples recovered were analyzed by quantitative PCR (qPCR) in the ABI 7500 real-time system. All experiments were performed in triplicate. The primer sequences used in the ChIP assay are as follows: HS3 forward primer,5′-ATAGACCATGAGTAGAGGGCAGAC-3′; HS3 reverse primer, 5′-TGATCCTGAAAACATAGGAGTCAA-3′; Aγ promoter forward primer, 5′-TTACTGCGCTGAAACTGTGG-3′; Aγ promoter reverse primer, 5′-CAGTGGTTTCTAAGGAAAAAGTGC-3′; Aγ (+3)kb forward primer, 5′-GAACAGAGCAGCACACTT-3′; Aγ (+3)kb reverse primer, 5′-TGAACAATAGTCCATGTCAAATCCT-3′; δ (−1)kb forward primer, 5′-GCAACAGAAGCCCAGCTATT-3′; δ (−1)kb reverse primer, 5′-GTGGCATGGTTTGATTTGTG-3′; β promoter forward primer, 5′-TGGTATGGGGCCAAGAGATA-3′; β promoter reverse primer, 5′-TGCTCCTGGGAGTAGATTGG-3′.

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Papers

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Manufacturer protocol

Download the product protocol from Abcam for Anti-Ctip1/BCL-11A antibody [14B5] (ab19487) below.

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Videos

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