Pan-methylarginine antibody generation using PEG linked GAR motifs as antigens

Arginine methylation of BRD4 by PRMT2/4 governs transcription and DNA repair

“…To test whether BRD4 is methylated by PRMT2/4, we performed Western blot using recently developed antibodies against MMA or ADMA ( 32 ). Overexpression of PRMT4 and, to a lesser extent, PRMT2, but not PRMT7, promoted MMA formation on BRD4 in an enzymatic-dependent manner ( Fig.…”

“…S2F). Notably, the ADMA signal was not detected using three widely used pan anti-ADMA antibodies, including anti-ADMA ( 32 ), adme-R (Cell Signaling Technology), and ASYM24 (Sigma-Aldrich). However, we could not rule out ADMA modification on BRD4 because it might not be recognized by these pan anti-ADMA antibodies.…”

Human protein arginine methyltransferases (PRMTs) can be optimally active under nonphysiological conditions

“…Such cross talk between PRMTs can complicate the analysis of specific PRMT activity in cells. Finally, it is now clear that reductions in the activity of one PRMT can result in the methylation of its sites by other members of the PRMT family ( 40 , 41 ). The identification of specific substrates and the generation of specific antibodies raised against their monomethylation site(s) will be important steps in quantitating PRMT7 activity in cells and testing how this activity may respond to changes in their intracellular environment.…”

PRMT blockade induces defective DNA replication stress response and synergizes with PARP inhibition

“…Tissues were washed in PBS-T (PBS +0.05% Triton X-100), blocked with 2% BSA in PBS-T, and then incubated overnight with 1:1000 anti-SDMA produced as previously described. 42 Tissues were washed, stained with anti-rabbit HRP (Cell Signaling), and Tyramide signal amplification (TSA) was performed with CF550R Dye (Biotium, #96077) in amplification buffer (Akoya Biosciences). Slides were subject to additional antigen retrieval in citrate buffer, and then the procedure repeated with anti-pan cytokeratin (clone AE1/3, Bethyl), anti-mouse HRP (Cell Signaling), and CF680R (Biotium, #92196).…”