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Phospho-POLR2A CTD-S2 Rabbit mAb (AP0996)

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, Mouse, Rat

Chromatin immunoprecipitations were performed with cross-linked chromatin from 293F cells and Phospho-POLR2A-S2 (AP0996). The ChIP sequencing results indicate the enrichment pattern of Phospho-POLR2A-S2 in selected genomic region and representative gene loci (GAPDH), as shown in figure.

ABclonal: - Phospho-POLR2A CTD-S2 Rabbit mAb (AP0996)

CUT&Tag was performed using the CUT&Tag Assay Kit(pAG-Tn5) forIllumina (RK20265) from 10⁵ Hela cells with 1μg Phospho-POLR2A CTD-S2 Rabbit mAb(AP0996), along with a Goat Anti-Rabbit IgG(H+L). The CUT&Tag results indicate the enrichment pattern of Phospho-POLR2A CTD-S2 in representative gene loci(GAPDH).

ABclonal:Western blot - Phospho-POLR2A CTD-S2 Rabbit mAb (AP0996)

Western blot analysis of various lysates using Phospho-POLR2A CTD-S2 Rabbit mAb (AP0996) at 1:1000 dilution. MCF7 cells and HeLa cells and C2C12 cells were treated by CIP(20uL/400ul) at 37℃ for 1 hour.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% BSA.
Detection: ECL Basic Kit (RM00020).
Exposure time: 10s.

Western blot analysis of lysates from C6 cells, using Phospho-POLR2A CTD-S2 Rabbit mAb (AP0996) at 1:1000 dilution. C6 cells were treated by CIP(20uL/400ul) at 37℃ for 1 hour.
Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) (AS014) at 1:10000 dilution.
Lysates/proteins: 25μg per lane.
Blocking buffer: 3% BSA.
Detection: ECL Basic Kit (RM00020).
Exposure time: 1min.

ABclonal:Immunohistochemistry - Phospho-POLR2A CTD-S2 Rabbit mAb (AP0996)

Immunohistochemistry analysis of paraffin-embedded Human cervix cancer using Phospho-POLR2A CTD-S2 Rabbit mAb (AP0996) at dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Mouse colon using Phospho-POLR2A CTD-S2 Rabbit mAb (AP0996) at dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.

Immunohistochemistry analysis of paraffin-embedded Rat kidney using Phospho-POLR2A CTD-S2 Rabbit mAb (AP0996) at dilution of 1:200 (40x lens). High pressure antigen retrieval performed with 0.01M Citrate Bufferr (pH 6.0) prior to IHC staining.

ABclonal:Immunofluorescence - Phospho-POLR2A CTD-S2 Rabbit mAb (AP0996)

Immunofluorescence analysis of A-549 cells using Phospho-POLR2A CTD-S2 Rabbit mAb (AP0996) at dilution of 1:100 (40x lens). Secondary antibody: Cy3-conjugated Goat anti-Rabbit IgG (H+L) (AS007) at 1:500 dilution. Blue: DAPI for nuclear staining.

Immunofluorescence analysis of HeLa cells using Phospho-POLR2A CTD-S2 Rabbit mAb (AP0996) at dilution of 1:100 (40x lens). Secondary antibody: Cy3-conjugated Goat anti-Rabbit IgG (H+L) (AS007) at 1:500 dilution. Blue: DAPI for nuclear staining.

Immunofluorescence analysis of NIH/3T3 cells using Phospho-POLR2A CTD-S2 Rabbit mAb (AP0996) at dilution of 1:100 (40x lens). Secondary antibody: Cy3-conjugated Goat anti-Rabbit IgG (H+L) (AS007) at 1:500 dilution. Blue: DAPI for nuclear staining.

Immunofluorescence analysis of PC-12 cells using Phospho-POLR2A CTD-S2 Rabbit mAb (AP0996) at dilution of 1:100 (40x lens). Secondary antibody: Cy3-conjugated Goat anti-Rabbit IgG (H+L) (AS007) at 1:500 dilution. Blue: DAPI for nuclear staining.

ABclonal:Chromatin Immunoprecipitation - Phospho-POLR2A CTD-S2 Rabbit mAb (AP0996)

Chromatin immunoprecipitation analysis of extracts of 293F cells, using Phospho-POLR2A-S2 antibody (AP0996) 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-POLR2A CTD-S2 Rabbit mAb
Catalog No.	AP0996
Host species	Rabbit
Purification method	Affinity purification
Isotype	IgG
CloneNo.	ARC1540

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 of human POLR2A CTD repeat YSPTSPS.
Sequence	Email for sequence
Gene ID
Swiss Prot
Synonyms	RPB1; RPO2; POLR2; POLRA; RPBh1; RPOL2; NEDHIB; RpIILS; hsRPB1; hRPB220; Phospho-POLR2A CTD-S2
Calculated MW	217kDa
Observed MW	270kDa

Applications

Reactivity	Human, Mouse, 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 1:500 - 1:2000 IHC-P 1:50 - 1:200 IF/ICC 1:50 - 1:200 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 ChIP-seq 1:50 - 1:100 CUT&Tag 10⁵ cells /1 μg
Storage buffer	Store at -20℃. Avoid freeze / thaw cycles. Buffer: PBS with 0.02% sodium azide, 0.05% BSA, 50% glycerol, pH7.3.
Key application	Western blotting Immunohistochemistry Immunofluorescence
Positive samples	MCF7, HeLa, C2C12, C6
Cellular location	Nucleus.
Customer validation	WB(Mus musculus)