Anti-Phospho-POLR2A CTD-S2 Polyclonal Antibody for WB, IP, ELISA - P24928

Core Product Specifications and Parameters

Biological Background: POLR2A (DNA-directed RNA polymerase II subunit RPB1) Function and Localization

• POLR2A encodes the largest and catalytic subunit of RNA polymerase II (Pol II), a DNA-dependent RNA polymerase responsible for synthesizing mRNA precursors and many functional non-coding RNAs (PubMed:23748380, PubMed:27193682, PubMed:30190596, PubMed:28108474). Related references: PMID:23748380 PMID:27193682 PMID:30190596 PMID:28108474
• Together with the second largest subunit POLR2B, it forms the Pol II active center, appending nucleotides to the nascent RNA and providing proofreading through a 3'->5' exonuclease activity (PubMed:30190596, PubMed:8381534, PubMed:9852112). Related references: PMID:30190596 PMID:8381534 PMID:9852112
• The unique C-terminal domain (CTD) consists of 52 tandem heptapeptide repeats (YSPTSPS) and serves as a dynamic platform for the assembly of factors regulating transcription initiation, elongation, termination, and co-transcriptional mRNA processing. Phosphorylation at Ser5 promotes promoter escape, while phosphorylation at Ser2 is critical for pause release and productive elongation (PubMed:24207025, PubMed:26124092). Related references: PMID:24207025 PMID:26124092
• In addition to DNA-dependent RNA polymerase activity, POLR2A possesses RNA-dependent RNA polymerase function, extending non-coding RNAs and participating in viral replication, e.g., with Hepatitis delta virus.
• Post-translational modifications of POLR2A include phosphorylation, acetylation, methylation, and ubiquitination, which finely tune its activity and interactions.
• POLR2A localizes predominantly to the nucleus but can also be found in the cytoplasm and associated with chromosomes, reflecting its roles in transcription and beyond.
• The calculated molecular weight is approximately 217 kDa.

Experimental Guidance and Technical Tips

• The immunogen is a synthetic phospho-peptide corresponding to Ser2 of the human POLR2A CTD repeat; antibody specificity should be confirmed using phosphatase-treated lysates or a non-phosphorylated blocking peptide.
• For Western blot applications, include appropriate phosphatase inhibitors in lysis and sample buffers to preserve the phosphorylation signal.
• When performing immunoprecipitation, ensure lysis buffer composition maintains phospho-epitope integrity; pre-clearing lysates may reduce nonspecific binding.
• For ELISA, coating with the immunizing phospho-peptide and detection with this antibody can be used to assess phospho-specific binding; consider cross-reactivity with unphosphorylated peptide controls.
• This antibody may also recognize other proteins with similar phospho-motifs; validate in the relevant sample system.

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