Products

ChIP assays were performed using human HeLa cells. A titration consisting of 1, 2, 5 and 10 ug of antibody per ChIP experiment was analyzed. IgG (2 ug/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter of the active genes c-fos and EIF2S3, used as positive controls, and for the inactive TSH2B gene and the Sat2 satellite repeat, used as negative controls.

ChIP assays were performed using human K562 cells. A titration of the antibody consisting of 0.2, 0.5, 1 and 2 ug per ChIP experiment was analysed. IgG (1 ug/IP) was used as negative IP control. Quantitative PCR was performed with primers specific for the promoter of the active genes c-fos and EIF2S3, used as positive controls, and for the coding region of the inactive MB gene and the Sat2 satellite repeat, used as negative controls. The figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

ChIP was performed on sheared chromatin from 100,000 K562 cells using 0.5 ug of antibody. The figure shows the peak distribution in four genomic regions including the regions surrounding the EIF2S3 and c-fos positive control genes on chromosome X and 14, respectively.

Western Blot analysis of (1) 25 ug whole cell extracts of Hela cells, (2) 15 ug histone extracts of Hela cells, (3) histone extracts after incubation of the antibody with 1 ug of the peptide used for immunisation of the rabbit, (4) histone extracts after incubation of the antibody with a peptide containing a sequence from the central part of the Histone H2AZ protein, (5) 1 ug of recombinant histone H2A, (6) 1 ug of recombinant histone H2B, (7) 1 ug of recombinant histone H3, (8) 1 ug of recombinant histone H4.

Immunofluorescent staining of Hela cell line with antibody followed by (A) an anti-rabbit antibody conjugated to Alexa488 (left). The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings (right). (B) After incubation of the antibody with 10 ng/ul of the peptide used for immunisation of the rabbit and (C) with a peptide containing a sequence from the central part of the Histone H2AZ protein.

ELISA is a quantitative method used to determine the titer of the antibody using a serial dilution of antibody against Histone H2AZ. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:87500.