Phospho-POLR2A CTD-S5 Rabbit mAb for WB, IHC-P, IF/ICC, ELISA - P24928

Core Product Specifications and Parameters

Biological Background: POLR2A (RPB1) Function and Localization

• POLR2A encodes DNA-directed RNA polymerase II subunit RPB1, the largest catalytic subunit of Pol II, also known as RPB1, hRPB220, and POLR2 (UniProt P24928).
• As the core catalytic component of Pol II, RPB1 synthesizes mRNA precursors and non-coding RNAs during the transcription cycle of initiation, elongation, and termination. Related references: PMID:23748380 PMID:27193682 PMID:30190596 PMID:28108474
• The C-terminal domain (CTD) consists of 52 heptapeptide repeats and undergoes dynamic phosphorylation: Ser5 phosphorylation promotes promoter escape, while Ser2 phosphorylation enables pause release and elongation. Related references: PMID:24207025 PMID:26124092
• RPB1 localizes predominantly to the nucleus, with additional presence in the cytoplasm and on chromosomes during transcription.
• Post-translational modifications of the CTD include acetylation, methylation, and ubiquitination, which modulate transcription initiation and elongation, especially of growth-factor-induced genes.
• RPB1 also possesses RNA-dependent RNA polymerase activity, implicated in non-coding RNA extension and host-virus interactions, such as Hepatitis delta virus replication.
• Mutations in POLR2A are linked to neurodevelopmental disorders and intellectual disability (UniProt keyword: Intellectual disability, Disease variant).

Experimental Guidance and Technical Tips

• The immunogen is a synthetic phospho-peptide corresponding to residues surrounding Ser5 of the POLR2A CTD, ensuring specificity for the phosphorylated form. Validate antibody specificity using phosphatase treatment or non-phosphorylated peptide blocking in your assay system.
• For Western blot, expect a band at ~217 kDa; consider using an appropriate gel percentage and transfer conditions for high molecular weight proteins.
• The antibody is validated for IHC-P and IF/ICC; optimize antigen retrieval and permeabilization conditions based on tissue or cell type.
• Cross-reactivity with mouse and rat suggests conserved sequences in these species; confirm suitability in your experimental model.

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