Phospho-POLR2A (Ser5) Recombinant monoclonal antibody

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  WB analysis using 84834-1-RR

  Non-treated HeLa cells, phosphatase inhibitor treated HeLa cells and λ phosphatase treated HeLa cells were subjected to SDS PAGE followed by western blot with 84834-1-RR (Phospho-POLR2A (Ser5) antibody) at dilution of 1:10000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with Lamin B1 (66095-1-Ig) antibody as a loading control.

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  IF Staining of HeLa using 84834-1-RR

  Immunofluorescent analysis of (-20°C Ethanol) fixed HeLa cells using Phospho-POLR2A (Ser5) antibody (84834-1-RR, Clone: 241914E5 ) at dilution of 1:400 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2).

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  FC experiment of HeLa using 84834-1-RR

  1X10^6 HeLa cells untreated (dashed lines) or treated with λ phosphatase which intracellularly stained with 0.13 ug Phospho-POLR2A (Ser5) Recombinant antibody (84834-1-RR, Clone:241914E5) and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2)(red), or 0.13 ug Rabbit IgG Isotype Control RecAb (98136-1-RR, Clone: 240953C9) (blue). Cells were fixed with 4% PFA and permeabilized with 90% MeOH.

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  ChIP experiment of HeLa using 84834-1-RR

  Chromatin was prepared from HeLa cells. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 20 µg of cross-linked chromatin, 5 µg of Phospho-POLR2A (Ser5) (84834-1-RR) or 5 ug of Normal Rabbit IgG (98136-1-RR), and 20 µl of Protein A Magarose Beads. The immunoprecipitated DNA was quantified by real-time PCR.

Phospho-POLR2A (Ser5) Uni-rAb^® Recombinant Antibody for WB, IF/ICC, FC (Intra), ELISA, ChIP-qPCR

Cat No. 84834-1-RR

Clone No.241914E5

Host / Isotype

Rabbit / IgG

Reactivity

human

Applications

WB, IF/ICC, FC (Intra), ELISA, ChIP-qPCR

Synonyms

Phospho POLR2A, Phospho POLR2A (Ser5), Phospho-POLR2A, phospho-polymerase II, Phospho-Rpb1 CTD

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Tested Applications

• Positive WB detected in: HeLa cells, λ phosphatase treated HeLa cells
• Positive IF/ICC detected in: HeLa cells
• Positive FC (Intra) detected in: λ phosphatase treated HeLa cells
• Positive ChIP-qPCR detected in: HeLa cells

Recommended dilution

• Western Blot (WB): WB : 1:5000-1:50000
• Immunofluorescence (IF)/ICC: IF/ICC : 1:200-1:800
• Flow Cytometry (FC) (INTRA): FC (INTRA) : 0.13 ug per 10^6 cells in a 100 µl suspension
• CHIP-QPCR: CHIP-QPCR : 1:10-1:100

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Product Information

84834-1-RR targets Phospho-POLR2A (Ser5) in WB, IF/ICC, FC (Intra), ELISA, ChIP-qPCR applications and shows reactivity with human samples.

• Tested Reactivity: human
• Host / Isotype: Rabbit / IgG
• Class: Recombinant
• Type: Antibody
• Immunogen:

  Peptide

• Full Name: polymerase (RNA) II (DNA directed) polypeptide A, 220kDa
• Calculated Molecular Weight: 1970 aa, 217 kDa
• Observed Molecular Weight: 270 kDa
• GenBank Accession Number: BC137231
• Gene Symbol: POLR2A
• Gene ID (NCBI): 5430
• RRID: AB_3672142
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purfication
• UNIPROT ID: P24928
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol, pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

RNA Pol II consists of 12 subunits and is responsible for the expression of mRNA and lincRNA genes within cells. The subunits of RNA polymerases have been shown to play specific roles in Pol II CTD dephosphorylation, tissue-specific transcription, mRNA decay, and translation. Pol II transcription involves initiation through recruitment and CTD serine 5 phosphorylation, elongation with CTD serine 2 phosphorylation, and termination. (PMID: 38842922)