Anti-POLR2A (N-terminal) Rabbit mAb for WB, IP, ChIP, ELISA - P24928

Core Product Specifications and Parameters

Biological Background: POLR2A Function and Localization

• POLR2A encodes the largest catalytic subunit of RNA polymerase II (Pol II), which synthesizes mRNA precursors and many functional non-coding RNAs using nucleoside triphosphates as substrates. Related references: PMID:23748380, PMID:27193682, PMID:30190596

• During transcription initiation, Pol II is recruited to DNA promoters, and POLR2A forms the active center with POLR2B, contributing a Mg²⁺-coordinating DxDGD motif essential for nucleotide addition. Related references: PMID:23748380, PMID:27193682, PMID:28108474, PMID:30190596, PMID:8381534, PMID:9852112

• The unique C-terminal domain (CTD) consists of 52 heptapeptide repeats and serves as a platform for assembly of factors regulating transcription initiation, elongation, termination, and co-transcriptional processing. Related references: PMID:24207025, PMID:26124092

• POLR2A exhibits intrinsic 3'→5' exonuclease proofreading activity, enabling removal of misincorporated nucleotides through backtracking and phosphodiester bond cleavage. Related references: PMID:8381534

• CTD phosphorylation on Ser-5 mediates promoter escape, while Ser-2 phosphorylation is required for pause release during elongation; methylation and acetylation of CTD lysines fine-tune gene expression levels. Related references: PMID:24207025, PMID:26124092

• Beyond DNA-dependent transcription, POLR2A can function as an RNA-dependent RNA polymerase, extending non-coding RNAs and replicating the hepatitis delta virus genome during infection. Related references: PMID:23748380, PMID:27193682, PMID:30190596

• The protein localizes to the nucleus, cytoplasm, and chromosomes, reflecting its dynamic roles in transcription and RNA processing.

• Alternative splicing generates multiple isoforms, and disease-associated variants in POLR2A have been linked to neurodevelopmental disorders (e.g., NEDHIB).

Experimental Guidance and Technical Tips

• The immunogen target (aa550-650) lies within the N-terminal region and is conserved across human, mouse, and rat, supporting the observed cross-reactivity; consider sequence alignment for other species.
• For Western blot, expect a band near 217 kDa; use a low-percentage (e.g., 6%) SDS-PAGE gel and optimize transfer conditions for high-molecular-weight proteins.
• In chromatin immunoprecipitation (ChIP), validate antibody specificity with positive and negative genomic loci, and optimize crosslinking and shearing protocols due to the large size of the Pol II complex.
• For immunoprecipitation (IP), mild lysis conditions and inclusion of nuclease treatment may improve solubility of chromatin-bound POLR2A.
• The antibody is also suitable for ELISA; consider pairing with a capture antibody for sandwich-based detection of native Pol II complexes.

CamelBio: Your One-Stop Sourcing Bridge

CamelBio streamlines IVD raw material procurement for diagnostic manufacturers and research laboratories, offering validated antibody pairs, optimized monoclonal and polyclonal antibodies, and ancillary bulk reagents. For transcription and RNA polymerase II research, this anti-POLR2A (N-terminal) rabbit mAb is part of a broader portfolio covering key Pol II subunits and transcription factors. Through its integrated supply network, CamelBio also assists with sourcing rare-target raw materials, helping customers accelerate development from concept to clinic.