Anti-POLR2A Rabbit Polyclonal Antibody for WB, IF/ICC, IP, ELISA - P24928

Core Product Specifications and Parameters

Biological Background: POLR2A Function and Localization

• POLR2A encodes the largest subunit (RPB1) of DNA-directed RNA polymerase II, the enzyme responsible for synthesizing mRNA precursors and many non-coding RNAs in eukaryotic cells.
• RPB1 forms the catalytic core of Pol II together with the second largest subunit RPB2. It contains a conserved DxDGD motif that coordinates a Mg²⁺ ion essential for nucleotide addition during RNA polymerization. Related references: PMID:30190596, PMID:8381534, PMID:9852112
• The enzyme cycles through transcription initiation, elongation, and termination. During initiation, Pol II is recruited to promoters containing elements like the TATA-box or CpG islands. Related references: PMID:23748380, PMID:27193682, PMID:28108474
• RPB1 possesses a unique C-terminal domain (CTD) composed of 52 heptapeptide repeats. The CTD undergoes dynamic phosphorylation (especially on Ser-2 and Ser-5) to coordinate transcription stages and recruit processing factors. Related references: PMID:24207025, PMID:26124092
• In addition to its DNA-dependent RNA polymerase activity, POLR2A can act as an RNA-dependent RNA polymerase, extending non-coding RNAs and mediating replication of hepatitis delta virus.
• The protein localizes predominantly to the nucleus, but is also found in the cytoplasm and associated with chromosomes during mitosis.
• Mutations in POLR2A are associated with neurodevelopmental disorders, including intellectual disability (NEDHIB), highlighting its essential role in gene expression.
• Alternative splicing and post-translational modifications (acetylation, methylation, ubiquitination, phosphorylation) further diversify POLR2A function and regulation.

Experimental Guidance and Technical Tips

• The immunogen corresponds to the N-terminal region (aa 1-260) of human POLR2A, which lies outside the repetitive CTD. This design may confer specificity for the RPB1 core over the highly homologous CTD found in some other proteins.
• The antibody has been demonstrated to work in WB, IF/ICC, IP, and ELISA. When performing WB, note the predicted molecular weight of ~217 kDa; consider using a low-percentage SDS-PAGE gel and thorough transfer optimization for such a large protein.
• Cross-reactivity with mouse and rat is reported, suggesting the epitope is conserved across these species. Validate in your specific model system to confirm.
• For IF/ICC, nuclear and perinuclear staining is expected based on POLR2A localization; however, background may vary depending on fixation and permeabilization conditions.
• In IP experiments, ensure the lysis buffer is compatible with nuclear protein extraction to effectively solubilize Pol II complexes.

CamelBio: Your One-Stop Sourcing Bridge

CamelBio supplies premium IVD raw materials and technical support to diagnostic developers and research laboratories worldwide, streamlining your journey from concept to clinic. This anti-POLR2A antibody serves as a versatile tool for studying RNA polymerase II and transcription regulation, complementing our portfolio of validated antibody pairs, bulk ancillary reagents, and customizable monoclonal/polyclonal antibody development services. Whether you need rare-target sourcing or ready-to-use critical raw materials, CamelBio is your trusted partner for accelerating assay and kit development.