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We have your antibodies against Histone H3K4 methylation (ab8895 for monomethylation and ab8580 for trimethylation). I would like to see their specificity on WB. Do you have any WB data loading Histon H3 unmodified, monomethylated H3K4, dimethyated H3K4 and trimethylated H3K4 and then stained by ab8895 or ab8580 ? Also do you have any control peptide/protein applicable for WB of H3K4 methylation WB ? They should be full size of monomethylated H3K4, dimethylated H3K4, trimethylated H3K4 and unmodified H3.
Asked on Feb 13 2012
Peptides are available for both of these products. The peptide for ab8895,Anti-Histone H3 (mono methyl K4) antibody - ChIP Grade, is product number ab1340 (https://www.abcam.com/histone-h3-peptide-mono-methyl-k4-ab1340.html).The peptide for ab8580,Anti-Histone H3 (tri methyl K4) antibody - ChIP Grade, is ab1342 (https://www.abcam.com/histone-h3-peptide-tri-methyl-k4-ab1342.html). These are the exact peptides used to create the antibodies. Each peptide used in the testing assay is available as well. (https://www.abcam.com/index.html?datasheet=1340,https://www.abcam.com/index.html?datasheet=7768),https://www.abcam.com/index.html?datasheet=1342,https://www.abcam.com/index.html?datasheet=1771,https://www.abcam.com/index.html?datasheet=1780,https://www.abcam.com/index.html?datasheet=2903.
These products are tested by peptide assay. Our data on western blot is displayed on the datasheet for each product. For ab8895 our test show itspecific for mono-methylated Lysine 4 of histone H3 and does not recognize di- or tri-methyl Lysine 4 nor methylation at Lysine 9. In the image on the datasheet, this is shown in lane 2where the activity of the antibody is specifically blocked by the addition of the immunizing peptide (ab1340) [https://www.abcam.com/Histone-H3-mono-methyl-K4-antibody-ChIP-Grade-ab8895.html#ab8895_6.gif].
Further,ab8580 are tested in Peptide Array against peptides to different Histone H3 modifications. Six dilutions of each peptide are printed on to the Peptide Array in triplicate and results are averaged before being plotted on to a graph. Results show strong binding to Histone H3 - tri methyl K4 peptide (ab1342), indicating that this antibody specifically recognises the Histone H3 - tri methyl K4 modification [https://www.abcam.com/Histone-H3-tri-methyl-K4-antibody-ChIP-Grade-ab8580.html#Histone-H3-Primary-antibodies-ab8580-153.jpg.]
Each of these products are thoroughly characterized and published. There are links to specific publications and th Abreviews from our customers on each datasheet.
Answered on Feb 13 2012