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Phospho-Rpb1 CTD-S2/S5 Rabbit mAb (AP1354)

Tested applications:WB IHC-P IF/ICC IP ChIP ChIP-seq RIP FC FC(Intra)ELISA MeDIP Nucleotide Array DB FACS CoIP CUT&Tag meRIP InhibitionReactivity:Human, Rat

CUT&Tag was performed using the CUT&Tag Assay Kit (pAG-Tn5) for Illumina(RK20265) from 10⁵ K562 cells with 1 μg Phospho-POLR2A-S2/S5 Rabbit mAb (AP1354), along with a Goat Anti-Rabbit IgG(H+L). The CUT&Tag results indicate the enrichment pattern of Phospho-POLR2A-S2/S5 in representative gene loci (RPL30), as shown in figure.

ABclonal:Western blot - Phospho-Rpb1 CTD-S2/S5 Rabbit mAb (AP1354)

Western blot analysis of various lysates, using Phospho-POLR2A-S2/S5 Rabbit mAb (AP1354) at 1:2000 dilution. HeLa cells were treated by λ-PP mixed solution (1ul) at 30℃ for 30 minutes.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL Basic Kit (RM00020).
Exposure time: 4s.

ABclonal:Immunoprecipitation - Phospho-Rpb1 CTD-S2/S5 Rabbit mAb (AP1354)

Immunoprecipitation analysis of 300 μg extracts of HeLa cells using 3 μg Phospho-POLR2A-S2/S5 Rabbit mAb (AP1354). Western blot was performed from the immunoprecipitate using Phospho-POLR2A-S2/S5 antibody (AP1354) at a dilution of 1:2000.

ABclonal:Chromatin Immunoprecipitation - Phospho-Rpb1 CTD-S2/S5 Rabbit mAb (AP1354)

Chromatin immunoprecipitation analysis of extracts of HeLa cells, using Phospho-POLR2A-S2/S5 Rabbit mAb (AP1354) and rabbit IgG.The amount of immunoprecipitated DNA was checked by quantitative PCR. Histogram was constructed by the ratios of the immunoprecipitated DNA to the input.

Overview

Product name	Phospho-Rpb1 CTD-S2/S5 Rabbit mAb
Catalog No.	AP1354
Host species	Rabbit
Purification method	Affinity purification
Isotype	IgG
CloneNo.	ARC54959

Background

This gene encodes the largest subunit of RNA polymerase II, the polymerase responsible for synthesizing messenger RNA in eukaryotes. The product of this gene contains a carboxy terminal domain composed of heptapeptide repeats that are essential for polymerase activity. These repeats contain serine and threonine residues that are phosphorylated in actively transcribing RNA polymerase. In addition, this subunit, in combination with several other polymerase subunits, forms the DNA binding domain of the polymerase, a groove in which the DNA template is transcribed into RNA.

Immunogen information

Immunogen	A phospho specific peptide corresponding to residues surrounding S2/S5 of human POLR2A.
Sequence	Email for sequence
Gene ID
Swiss Prot
Synonyms	RPB1; RPO2; POLR2; POLRA; RPBh1; RPOL2; NEDHIB; RpIILS; hsRPB1; hRPB220; Phospho-POLR2A-S2/S5
Calculated MW	217kDa
Observed MW	270kDa

Applications

Reactivity	Human, Rat
Tested applications	WB IHC-P IF/ICC IP ChIP ChIP-seq RIP FC FC(Intra)ELISA MeDIP Nucleotide Array DB FACS CoIP CUT&Tag meRIP Inhibition
Recommended dilution	WB 1:1000 - 1:5000 IP 0.5μg-4μg antibody for 200μg-400μg extracts of whole cells ELISA Recommended starting concentration is 1 μg/mL. Please optimize the concentration based on your specific assay requirements. ChIP 5μg antibody for 10μg-15μg of Chromatin CUT&Tag 10⁵ cells /1 μg
Storage buffer	Store at -20℃. Avoid freeze / thaw cycles. Buffer: PBS with 0.05% proclin300, 0.05% BSA, 50% glycerol, pH7.3.
Key application	Western blotting Immunoprecipitation
Positive samples	HeLa
Cellular location	Nucleus.