WB
Western blot analysis of Phospho-POLR2A (S2) on different lysates with Rabbit anti-Phospho-POLR2A (S2) antibody at 1/1000 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. Exposure time: 6 seconds; 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 at 1/1000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50000 dilution was used for 1 hour at room temperature.ICC/IF
Immunocytochemistry analysis of HeLa cells labeling Phospho-POLR2A (S2) with Rabbit anti-Phospho-POLR2A (S2) antibody at 1/100 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 at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1000 dilution.ICC/IF
Immunocytochemistry analysis of RAW264.7 cells labeling Phospho-POLR2A (S2) with Rabbit anti-Phospho-POLR2A (S2) antibody 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 at 1/50 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L ( 594) was used as the secondary antibody at 1/1000 dilution.ICC/IF
Immunocytochemistry analysis of PC-12 cells labeling Phospho-POLR2A (S2) with Rabbit anti-Phospho-POLR2A (S2) antibody at 1/100 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 at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.IHC
Immunohistochemical analysis of paraffin-embedded human testis tissue untreated / treated with λpp with Rabbit anti-Phospho-POLR2A (S2) antibody at 1/1000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1000 dilution for 1 hour 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.IHC
Immunohistochemical analysis of paraffin-embedded mouse testis tissue untreated / treated with λpp with Rabbit anti-Phospho-POLR2A (S2) antibody at 1/1000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1000 dilution for 1 hour 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.IHC
Immunohistochemical analysis of paraffin-embedded rat testis tissue untreated / treated with λpp with Rabbit anti-Phospho-POLR2A (S2) antibody at 1/1000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1000 dilution for 1 hour 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.FC
Flow cytometric analysis of Phospho-POLR2A (S2) was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (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 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Product Name | Phospho-POLR2A (S2) Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Immunogen | Synthetic phospho-peptide corresponding to residues surrounding Ser2 of Human POLR2A aa 1590-1630 / 1970. |
| Modification | P-S2 |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIHCWB |
| WB:1:500-1:2000 ICC/IF:1:50-1:100 IHC:1:1000 FC:1:1000 |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Affinity purified |
| Gene Symbol | POLR2A |
|---|---|
| Gene Synonyms | RPB1 RPO2 POLR2 POLRA RPBh1 RPOL2 NEDHIB RpIILS hsRPB1 hRPB220 |
| Gene Full Name | RNA polymerase II subunit A |
| Gene Summary | 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. [provided by RefSeq, Jul 2008] |
| Alternative Names | DNA directed RNA polymerase II A antibody DNA-directed RNA polymerase II largest subunit RNA polymerase II 220 kd subunit antibody DNA-directed RNA polymerase II subunit A antibody DNA-directed RNA polymerase II subunit RPB1 antibody DNA-directed RNA polymerase III largest subunit antibody hRPB220 antibody hsRPB1 antibody POLR2 antibody Polr2a antibody POLRA antibody DNA directed RNA polymerase II A antibody DNA-directed RNA polymerase II largest subunit RNA polymerase II 220 kd subunit antibody DNA-directed RNA polymerase II subunit A antibody DNA-directed RNA polymerase II subunit RPB1 antibody DNA-directed RNA polymerase III largest subunit antibody hRPB220 antibody hsRPB1 antibody POLR2 antibody Polr2a antibody POLRA antibody Polymerase (RNA) II (DNA directed) polypeptide A 220kDa antibody Polymerase (RNA) II (DNA directed) polypeptide A antibody RNA polymerase II subunit B1 antibody RNA-directed RNA polymerase II subunit RPB1 antibody RPB1 antibody RPB1_HUMAN antibody RPBh1 antibody RpIILS antibody RPO2 antibody RPOL2 antibody |
| Molecular Weight(MW) | 217kDa(Observed band size:260kDa) |
| Cellular Localization | Nucleus, Cytoplasm, Chromosome. |
WB
Western blot analysis of Phospho-POLR2A (S2) on different lysates with Rabbit anti-Phospho-POLR2A (S2) antibody at 1/1000 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. Exposure time: 6 seconds; 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 at 1/1000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50000 dilution was used for 1 hour at room temperature.
ICC/IF
Immunocytochemistry analysis of HeLa cells labeling Phospho-POLR2A (S2) with Rabbit anti-Phospho-POLR2A (S2) antibody at 1/100 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 at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1000 dilution.
ICC/IF
Immunocytochemistry analysis of RAW264.7 cells labeling Phospho-POLR2A (S2) with Rabbit anti-Phospho-POLR2A (S2) antibody 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 at 1/50 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L ( 594) was used as the secondary antibody at 1/1000 dilution.
ICC/IF
Immunocytochemistry analysis of PC-12 cells labeling Phospho-POLR2A (S2) with Rabbit anti-Phospho-POLR2A (S2) antibody at 1/100 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 at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.
IHC
Immunohistochemical analysis of paraffin-embedded human testis tissue untreated / treated with λpp with Rabbit anti-Phospho-POLR2A (S2) antibody at 1/1000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1000 dilution for 1 hour 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.
IHC
Immunohistochemical analysis of paraffin-embedded mouse testis tissue untreated / treated with λpp with Rabbit anti-Phospho-POLR2A (S2) antibody at 1/1000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1000 dilution for 1 hour 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.
IHC
Immunohistochemical analysis of paraffin-embedded rat testis tissue untreated / treated with λpp with Rabbit anti-Phospho-POLR2A (S2) antibody at 1/1000 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/1000 dilution for 1 hour 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.
FC
Flow cytometric analysis of Phospho-POLR2A (S2) was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (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 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).| Application Notes | WB:1:500-1:2000 ICC/IF:1:50-1:100 IHC:1:1000 FC:1:1000 |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | 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), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
