Phospho-POLR2A (S5) Recombinant Rabbit Monoclonal Antibody [JM51-21]
cat.: ET1703-87
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: JM51-21
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: 250 kDa
Isotype: IgG
Immunogen: Synthetic phospho-peptide corresponding to residues surrounding Ser5 of Human POLR2A aa 1590-1627 / 1970.
Positive control: Hela, MCF-7, PC-12, human breast carcinoma tissue, human spleen tissue, human kidney tissue, mouse bladder 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
ET1703-87_1.jpg Fig1: Western blot analysis of Phospho-POLR2A (S5) on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1703-87, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.
Positive control:
Line 1: Hela cell lysate
Line 2: MCF-7 cell lysate
ET1703-87_2.jpg Fig2: ICC staining of Phospho-POLR2A (S5) 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 (ET1703-87, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1703-87_3.jpg Fig3: ICC staining of Phospho-POLR2A (S5) 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 (ET1703-87, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1703-87_4.jpg Fig4: ICC staining of Phospho-POLR2A (S5) 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 (ET1703-87, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
ET1703-87_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-Phospho-POLR2A (S5) 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 (ET1703-87, 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.
ET1703-87_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-Phospho-POLR2A (S5) 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 (ET1703-87, 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.
ET1703-87_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-Phospho-POLR2A (S5) 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 (ET1703-87, 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.
ET1703-87_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded mouse bladder tissue using anti-Phospho-POLR2A (S5) 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 (ET1703-87, 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.
ET1703-87_9.jpg Fig9: Flow cytometric analysis of Phospho-POLR2A (S5) was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1703-87, 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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