POLR2A Mouse Monoclonal Antibody [A5F3]
cat.: HA600062
Product Type: Mouse monoclonal IgG2b, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IHC-P
Clonality: Monoclonal
Clone number: A5F3
Form: Liquid
Storage condition: Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
Storage buffer: 1*TBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 2ug/ul
Purification: Protein G affinity purified.
Molecular weight: Predicted band size: 217 kDa
Isotype: IgG2b
Immunogen: Recombinant protein within human POLR2A aa 751-950 / 1970.
Positive control: HeLa cell lysate, MCF7 cell lysate, 293T cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, HeLa, rat bladder tissue, human placenta tissue, mouse testis tissue.
Subcellular location: Cytoplasm, Nucleus, Chromosome.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
1:2,000
1:50-1:100
1:100-1:500
Uniprot #: SwissProt: P24928 Human | P08775 Mouse | D4A5A6 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
HA600062_1.jpg Fig1: Western blot analysis of POLR2A on different lysates with Mouse anti-POLR2A antibody (HA600062) at 1/2,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: MCF7 cell lysate
Lane 3: 293T cell lysate
Lane 4: NIH/3T3 cell lysate
Lane 5: PC-12 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 217 kDa
Observed band size: 250 kDa

Exposure time: 4 seconds; ECL: K1801;

3-8% 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 (HA600062) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.
HA600062_2.jpg Fig2: Immunocytochemistry analysis of HeLa cells labeling POLR2A with Mouse anti-POLR2A antibody (HA600062) 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 Mouse anti-POLR2A antibody (HA600062) at 1/50 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) 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 (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.
HA600062_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded rat bladder tissue using anti-POLR2A antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (HA600062, 1/400) 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.
HA600062_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human placenta tissue using anti-POLR2A antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (HA600062, 1/100) 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.
HA600062_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded mouse testis tissue using anti-POLR2A antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 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 (HA600062, 1/400) 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.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.