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.
Concentration: 100 µl Formulation: Whole antiserum from rabbit containing 0.05% azide Species Reactivity: Human Purification: Whole serum Immunogen: synthetic peptide Format: aqueous solution Storage / Stability: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.
Application(s): ELISA (1:100 – 1:500) DB (1:1000) WB (1:250) Notes: The optimal antibody concentration should be determined by the end–user. Warning(s): Avoid freeze/thaw cycles Scientific Category: Antibodies Product Type: Antibody Data shown is lot-specific. Contact us for specific information on other lots.
