![Western Blot: RNA Polymerase II/POLR2A Antibody (4H8) [NB200-598] - Whole cell protein from HeLa (lane 1), MCF7 (lane 2), NIH-3T3 (lane 3) and Cos7 (lane 4) were separated by SDS-PAGE on a 6% gel, transferred to PVDF and probed with 2 ug/mL NB200-598. RNA Pol II reactivity (arrowhead) is observed in HeLa, MCF and 3T3 cells.](http://images.novusbio.com/fullsize/RNA-Polymerase-II-POLR2A-Antibody-4H8-Western-Blot-NB200-598-img0010.jpg "Western Blot: RNA Polymerase II/POLR2A Antibody (4H8) [NB200-598] - Whole cell protein from HeLa (lane 1), MCF7 (lane 2), NIH-3T3 (lane 3) and Cos7 (lane 4) were separated by SDS-PAGE on a 6% gel, transferred to PVDF and probed with 2 ug/mL NB200-598. RNA Pol II reactivity (arrowhead) is observed in HeLa, MCF and 3T3 cells.")
| Reactivity | Hu, Mu, Ye, Pm(-), Dr, Ha, PlSpecies Glossary |
| Applications | WB, ELISA, Flow, ICC/IF, IP, ChIP, CyTOF-ready |
| Clone | 4H8 |
| Clonality | Monoclonal |
| Host | Mouse |
| Conjugate | Unconjugated |
| Format | Azide and BSA Free |
| Concentration | 1 mg/ml |
| Immunogen | 10 repeats of synthetic peptide YSPTSPS using chemically synthesized phospho-Ser 5 YSPTSpPS (Human). [UniProt# P24928] |
| Specificity | Please note that this antibody detects both unphosphorylated and phosphorylated forms of the protein. When using this antibody in western blot and IP, it recognizes both the II0 and IIA form of RPB1 (ie the largest RNAPII subunit). These different forms of RPB1 (as seen in gels) have traditionally been loosely defined as 'hyper-' and 'hypo-phosphorylated', respectively. Here, however, it is important to remember that RPB1 contains no less than 52 repeats of a heptapeptide (TSPTSPS) that can be phosphorylated at different positions (Tyr1, Ser2, Thr4, Ser5 and Ser7; this antibody was raised against a synthetic peptide with 10 of these repeats, mostly but probably not entirely, chemically phosphorylated at Ser5). So, when we say that it recognizes both forms (II0/hyper-phos and IIA/hypo-phos), it might really just mean that the antibody recognizes a few phosphorylated Ser5s among 52 otherwise mostly un-phosphorylated repeats. Alternatively, it might conceivably (though less likely) recognize UN-phosphorylated repeats as there might conceivably be a few of those even in the II0/hyper-phos RPB1 form.' |
| Predicted Species | Plant (100%), Drosophila (100%), Hamster (100%). Backed by our 100% Guarantee. |
| Isotype | IgG1 |
| Clonality | Monoclonal |
| Host | Mouse |
| Gene | POLR2A |
| Purity | Protein G purified |
| Innovator's Reward | Test in a species/application not listed above to receive a full credit towards a future purchase. |
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| Application Notes | In WB a band can be seen at approximately 217 kDa. Optimal dilutions/concentrations should be determined by the end user. |
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| Theoretical MW | 217 kDa. Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors. |
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| Storage | Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles. |
| Buffer | PBS |
| Preservative | No Preservative |
| Concentration | 1 mg/ml |
| Purity | Protein G purified |
| Images | Ratings | Applications | Species | Date | Details | ||||
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reviewed by:
Bryan Tinsley |
WB | Human | 05/13/2014 |
Summary
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Secondary Antibodies |
Isotype Controls |
Research Areas for RNA Polymerase II/POLR2A Antibody (NB200-598)Find related products by research area.
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![Immunocytochemistry/Immunofluorescence: RNA Polymerase II/POLR2A Antibody (4H8) [NB200-598] - RNA Polymerase II/POLR2A antibody was tested in HeLa cells with DyLight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and DyLight 550 (red).](http://images.novusbio.com/fullsize/RNA-Polymerase-II-POLR2A-Antibody-4H8-Immunocytochemistry-Immunofluorescence-NB200-598-img0011.jpg "Immunocytochemistry/Immunofluorescence: RNA Polymerase II/POLR2A Antibody (4H8) [NB200-598] - RNA Polymerase II/POLR2A antibody was tested in HeLa cells with DyLight 488 (green). Nuclei and alpha-tubulin were counterstained with DAPI (blue) and DyLight 550 (red).")
![Flow Cytometry: RNA Polymerase II/POLR2A Antibody (4H8) [NB200-598] - An intracellular stain was performed on HeLa cells with RNA Polymerase II/POLR2A (4H8) antibody NB200-598PE (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes. Both antibodies were conjugated to phycoerythrin.](http://images.novusbio.com/fullsize/RNA-Polymerase-II-POLR2A-Antibody-4H8-Flow-Cytometry-NB200-598-img0013.jpg "Flow Cytometry: RNA Polymerase II/POLR2A Antibody (4H8) [NB200-598] - An intracellular stain was performed on HeLa cells with RNA Polymerase II/POLR2A (4H8) antibody NB200-598PE (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 2.5 ug/mL for 30 minutes. Both antibodies were conjugated to phycoerythrin.")
![Western Blot: RNA Polymerase II/POLR2A Antibody (4H8) [NB200-598] - Analysis of MCF7 cell lysata (A) using RNA polymerase II antibody at 2 ug/mL.](http://images.novusbio.com/fullsize/RNA-Polymerase-II-POLR2A-Antibody-4H8-Western-Blot-NB200-598-img0009.jpg "Western Blot: RNA Polymerase II/POLR2A Antibody (4H8) [NB200-598] - Analysis of MCF7 cell lysata (A) using RNA polymerase II antibody at 2 ug/mL.")
![Flow (Intracellular): RNA Polymerase II/POLR2A Antibody (4H8) [NB200-598] - An intracellular stain was performed on Daudi cells with RNA Polymerase II/POLR2A (4H8) antibody NB200-598 (blue) and a matched isotype control NBP2-27287 (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes, followed by mouse F(ab)2 IgG (H+L) APC-conjugated secondary antibody (F0101B, R&D Systems).](http://images.novusbio.com/fullsize/RNA-Polymerase-II-POLR2A-Antibody-4H8-Flow-Intracellular-NB200-598-img0012.jpg "Flow (Intracellular): RNA Polymerase II/POLR2A Antibody (4H8) [NB200-598] - An intracellular stain was performed on Daudi cells with RNA Polymerase II/POLR2A (4H8) antibody NB200-598 (blue) and a matched isotype control NBP2-27287 (orange). Cells were fixed with 4% PFA and then permeablized with 0.1% saponin. Cells were incubated in an antibody dilution of 1 ug/mL for 30 minutes, followed by mouse F(ab)2 IgG (H+L) APC-conjugated secondary antibody (F0101B, R&D Systems).")
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![SDS-Page: Mouse IgG1 Isotype Control (MG1) [NBP1-97005] - Lane 1: , Non-reduced. M: Opal Pre-stained Ladder. Lane 2: , Reduced. Load: 1.0 ug per lane. Predicted/Observed: 120 kDa Non-reduced, 55 and 25 Reduced.](https://images.novusbio.com/images/Mouse-IgG1-Isotype-Control-MG1-SDS-Page-NBP1-97005-img0002.jpg "SDS-Page: Mouse IgG1 Isotype Control (MG1) [NBP1-97005] - Lane 1: , Non-reduced. M: Opal Pre-stained Ladder. Lane 2: , Reduced. Load: 1.0 ug per lane. Predicted/Observed: 120 kDa Non-reduced, 55 and 25 Reduced.")
-(01-ml)_NB200-598_7561.jpg)