Phospho-POLR2A (S2) Recombinant Rabbit Monoclonal Antibody [JM11-51]
cat.: ET1703-86
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IP, IF-Cell, IF-Tissue, IHC-P, FC
Clonality: Monoclonal
Clone number: JM11-51
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 217 kDa
Isotype: IgG
Immunogen: Synthetic phospho-peptide corresponding to residues surrounding Ser2 of Human POLR2A aa 1590-1630 / 1970.
Positive control: HeLa cell lysate, RAW264.7 cell lysate, PC-12 cell lysate, RAW264.7, Hela, MCF-7, PC-12, human tonsil tissue, rat brain tissue, mouse testis tissue.
Subcellular location: Nucleus.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  FC
1:500-1:2,000
1:100-1:500
1:100-1:500
1:50-1:200
1:50-1:100
Uniprot #: SwissProt: P24928 Human | P08775 Mouse
Entrez Gene: 363633 Rat
Alternative names: DNA directed RNA polymerase II A DNA-directed RNA polymerase II largest subunit RNA polymerase II 220 kd subunit DNA-directed RNA polymerase II subunit A DNA-directed RNA polymerase II subunit RPB1 DNA-directed RNA polymerase III largest subunit hRPB220 hsRPB1 POLR2 Polr2a POLRA Polymerase (RNA) II (DNA directed) polypeptide A 220kDa Polymerase (RNA) II (DNA directed) polypeptide A RNA polymerase II subunit B1 RNA-directed RNA polymerase II subunit RPB1 RPB1 RPB1_HUMAN RPBh1 RpIILS RPO2 RPOL2
Images
ET1703-86_1.jpg Fig1: Western blot analysis of Phospho-POLR2A (S2) on different lysates with Rabbit anti-Phospho-POLR2A (S2) antibody (ET1703-86) at 1/1,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: RAW264.7 cell lysate
Lane 3: PC-12 cell lysate
Lane 4: HeLa cell lysate, the membrane treated with λpp for 1 hour

Lysates/proteins at 20 µg/Lane.

Predicted band size: 217 kDa
Observed band size: 260 kDa

Exposure time: 6 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1703-86) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
ET1703-86_2.jpg Fig2: Immunocytochemistry analysis of RAW264.7 cells labeling Phospho-POLR2A (S2) with Rabbit anti-Phospho-POLR2A (S2) antibody (ET1703-86) at 1/50 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Phospho-POLR2A (S2) antibody (ET1703-86) at 1/50 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
ET1703-86_3.jpg Fig3: ICC staining of Phospho-POLR2A (S2) in Hela cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1703-86, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1703-86_4.jpg Fig4: ICC staining of Phospho-POLR2A (S2) in MCF-7 cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1703-86, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1703-86_5.jpg Fig5: ICC staining of Phospho-POLR2A (S2) in PC-12 cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1703-86, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1703-86_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-Phospho-POLR2A (S2) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1703-86, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1703-86_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded rat brain tissue using anti-Phospho-POLR2A (S2) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1703-86, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1703-86_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded mouse testis tissue using anti-Phospho-POLR2A (S2) antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 20 minutes. The tissues were blocked in 1% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1703-86, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
ET1703-86_9.jpg Fig9: Flow cytometric analysis of Phospho-POLR2A (S2) was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1703-86, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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