Generation and Characterization of ADMA-Targeting Antibodies

D4H5, D6A8 and D10F7 were generated against degenerate peptides containing four asymmetric dimethylarginine (ADMA) residues XXXXRme2aXXRme2aXXXXRme2aXXX Rme2aX by Cell Signaling Technology (CST). D4H5 and D6A8 have been previously described20 (link) and are commercially available as part of an ADMA antibody mixture (CST, Catalog #13522). Anti CAS3 antibodies were custom raised by Bethyl laboratories against a peptide pool comprising amino acids 1-20 and 176-195 of human CAS3. CAS3 is also known asFAM168B or MANI; NM_001009993). Anti-methyl CAS3R87me2a (meCAS3) was generated against the epitope CTAVYPVRme2aSAYPQQSR by Covance. Methyl MED12R1899me2a (meMED12) was custom generated against the epitope TSVYRme2aQQQP by CST. PRMT1 antibody was a gift from Stephane Richard (McGill University). H3R2me2a (Millipore, Catalog #05-808), H3R17me2a (Millipore, Catalog #07-214), H3R8me2a (Active Motif, Catalog #39651), CARM1 (Bethyl, Catalog #A300-421A) and β-actin (Sigma, Catalog #A5316) were obtained commercially.

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Gayatri S., Cowles M.W., Vemulapalli V., Cheng D., Sun Z.W, & Bedford M.T. (2016). Using oriented peptide array libraries to evaluate methylarginine-specific antibodies and arginine methyltransferase substrate motifs. Scientific Reports, 6, 28718.

Publication 2016

H3R17me2a CARM1 β-actin

Actin Amino acids Anti antibodies Antibody Asymmetric dimethylarginine Carm1 Epitope Human Peptide

Corresponding Organization :

Other organizations : The University of Texas MD Anderson Cancer Center

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Variable analysis

independent variables

Degenerate peptides containing four asymmetric dimethylarginine (ADMA) residues: XXXXRme2aXXRme2aXXXXRme2aXXXRme2aX

dependent variables

Antibodies generated against the degenerate peptides: D4H5, D6A8, and D10F7
Anti CAS3 antibodies custom raised against a peptide pool comprising amino acids 1-20 and 176-195 of human CAS3
Anti-methyl CAS3R87me2a (meCAS3) antibody generated against the epitope CTAVYPVRme2aSAYPQQSR
Methyl MED12R1899me2a (meMED12) antibody generated against the epitope TSVYRme2aQQQP

control variables

H3R2me2a (Millipore, Catalog #05-808), H3R17me2a (Millipore, Catalog #07-214), H3R8me2a (Active Motif, Catalog #39651), CARM1 (Bethyl, Catalog #A300-421A) and β-actin (Sigma, Catalog #A5316) were obtained commercially

controls

Positive controls: Not explicitly mentioned
Negative controls: Not explicitly mentioned

Annotations

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