Polyclonal antibody raised in rabbit against histone H3 containing the acetylated lysine 23 (H3K23ac), using a KLH–conjugated synthetic peptide.

Determination of the titer
To determine the titer, an ELISA was performed using a serial dilution of the antibody directed against H3K23ac (Cat. No. 25237). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution (Figure 1), the titer of the antibody was estimated to be 1:2,750.

Cross reactivity test using the antibody directed against H3K23ac
A Dot Blot analysis was performed to test the cross reactivity of the antibody against H3K23ac (Cat. No. 25237) with with peptides containing other modifications and unmodified sequences of histone H3. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:1,000. Figure 2 shows a high specificity of the antibody for the modification of interest.

Western blot analysis using the antibody against H3K23ac
HeLa cells were treated with butyrate and histone extracts (15 μg) were analysed by Western blot using the antibody against H3K23ac (Cat. No. 25237) diluted 1:250 in TBS-Tween containing 5% skimmed milk (lane 1). Lane 2 shows a coomassie blue staining of the gel. The position of the protein of interest is indicated on the left; the marker (in kDa) is shown on the right.

100 µl

Whole antiserum from rabbit containing 0.05% azide

Human

Whole serum

synthetic peptide

aqueous solution

Store at –80°C for up to 2 years. Centrifuge after first thaw to maximize product recovery. Aliquot to avoid repeated freeze/thaw cycles. Aliquots may be stored at –20°C for at least one month.

ELISA (1:100 – 1:500)
DB (1:1000)
WB (1:250)

The optimal antibody concentration should be determined by the end–user.

Avoid freeze/thaw cycles

Antibodies

Antibody