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Purified anti-RNA Polymerase II RPB1 Antibody

Pricing & Availability
Clone
8WG16 (See other available formats)
Regulatory Status
RUO
Other Names
RNA polymerase II sub-unit 1, RPB1, CTD repeat YSPTSPS, C- terminal domain repeat YSPTSPS, DNA-directed RNA polymerase II subunit A, RNA-directed RNA polymerase II subunit RPB1, POLR2A, POLR2
Isotype
Mouse IgG2a
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Product Citations
publications
8WG16_Purified_RPB1_Antibody_1_WB_082316
Total lysates (15 µg protein) from HeLa cells were resolved by electrophoresis (4-12% Bis-Tris gel), transferred to nitrocellulose, and probed with 1:500 diluted (1 µg/mL) Purified anti-RNA Polymerase II RBP1 Antibody, clone 8WG16 (upper) or 1:3000 diluted Purified anti-β-actin Antibody, clone Poly6221 (lower). Proteins were visualized by chemiluminescence detection using a 1:3000 diluted goat anti-mouse-IgG secondary antibody conjugated to HRP for the anti-RNA Polymerase II RBP1 Antibody, and a donkey anti-rabbit IgG Antibody conjugated to HRP for anti-β-actin Antibody.
  • 8WG16_Purified_RPB1_Antibody_1_WB_082316
    Total lysates (15 µg protein) from HeLa cells were resolved by electrophoresis (4-12% Bis-Tris gel), transferred to nitrocellulose, and probed with 1:500 diluted (1 µg/mL) Purified anti-RNA Polymerase II RBP1 Antibody, clone 8WG16 (upper) or 1:3000 diluted Purified anti-β-actin Antibody, clone Poly6221 (lower). Proteins were visualized by chemiluminescence detection using a 1:3000 diluted goat anti-mouse-IgG secondary antibody conjugated to HRP for the anti-RNA Polymerase II RBP1 Antibody, and a donkey anti-rabbit IgG Antibody conjugated to HRP for anti-β-actin Antibody.
  • 8WG16_Purified_RPB1_Antibody_2_WB_022315
    Total cell lysates (15 µg protein) from HeLa, THP-1, and Raw264.7 were resolved by 4-12% Bis-tris gel electrophoresis, transferred to nitrocellulose, and probed with anti-RNA Polymerase II RPB1 antibody (clone 8WG16). Proteins were visualized using a goeat anti-mouse IgG secondary antibody conjugated to HRP and chemiluminescence detection. Direct-Blot™ HRP anti-β-actin antibody was used as a loading control.
  • 8WG16_Purified_RPB1_Antibody_3_ICC_042419
    HeLa cells were fixed with 2% paraformaldehyde (PFA) for ten minutes, permeabilized with 0.5% Triton X-100 for five minutes, and blocked with 5% FBS for 30 minutes. Then the cells were intracellularly stained with 2 µg/ml anti-RNA Polymerase II RPB1 (clone 8WG16 ) in blocking buffer overnight at 4°C and followed by DyLight™ 594 (red) conjugated goat anti-mouse IgG for one hour at room temperature. Actin filaments were labeled with Alexa Fluor® 488 Phalloidin (green). Nuclei were counterstained with DAPI (blue). The image was captured with a 60X objective.
  • 9-8WG16_GoChIP_RNAPolymeraseII_Antibody_050918
    Chromatin Immunoprecipitations (ChIP) were performed with cross-linked chromatin samples from 4 X 106 of HeLa cells with either A)1:300 dilution of Go-ChIP-Grade™ Purified anti-RNA Polymerase II RPB1 Antibody (clone 8WG16, Cat. No. 664911) or B) equal amount of Purified Mouse IgG2a, κ Isotype Control Antibody (Clone MOPC-173, Cat. No. 400201) by using Go-ChIP-Grade™ Protein G Enzymatic Kit (Cat. No. 699904). The enriched DNA was purified and quantified by real-time qPCR using primers targeting human GAPDH gene region and α-Satellite repeats. The amount of immunoprecipitated DNA in each sample is represented as signal relative to total amount of input chromatin.
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664906 100 µg 323 CHF
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Description

RPB1 is the catalytic and largest component of RNA polymerase II, which synthesizes mRNA precursors and many functional non-coding RNAs. It forms the polymerase active center together with RPB2, the second largest subunit. Polymerase II (Pol II) is the central component of the basal RNA polymerase II transcription machinery. It is composed of mobile elements that move relatively to each other. RPB1 is part of the core element with the central large cleft, the clamp element that moves to open and close the cleft, and the jaws that are thought to grab the incoming DNA template. At the start of transcription, a single DNA template strand of the promoter is positioned within the central active site cleft of Pol II. Then, a bridging helix emanates from RPB1 and crosses the cleft near the catalytic site, which acts as a ratchet that moves the RNA-DNA hybrid through the active site by switching from straight to bent conformations during each neuocleotide addition. This promotes translocation of Pol II. Pol II moves on the template during transcription elongation. Elongation is influenced by the phosphorylation status of the C-terminal domain (CTD) of Pol II's largest subunit (RPB1), which serves as a platform for assembling factors that regulate transcription initiation, elongation, termination, and mRNA processing. It can act as a RNA-dependent RNA polymerase when associated with small delta antigen of Hepatitis delta virus, being able to conform as both a replicate and transcriptase for the viral RNA circular genome.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human, Mouse
Reported Reactivity
Other species
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
Wheat germ RNA Polymerase II
Formulation
Phosphate-buffered solution, pH 7.2.
Preparation
The antibody was purified by affinity chromatography.
Concentration
1.0 mg/mL
Storage & Handling
This antibody should be handled aseptically as it is free of preservatives such as Sodium Azide. Store this antibody undiluted between 2°C and 8°C. Please note the storage condition for this antibody has been changed from -20°C to between 2°C and 8°C. You can also check the vial label or CoA to find the proper storage conditions.
Application

WB - Quality tested
ICC, ChIP - Verified
FA, IP - Reported in the literature, not verified in house
Recommended Usage

Each lot of this antibody is quality control tested by Western blotting. For Western blotting, the suggested use of this reagent is 0.25 - 2.0 µg per mL. For immunocytochemistry, a concentration range of 1.0 - 5.0 μg/mL is recommended. The suggested dilution for ChIP application is 1:300 - 1:500 by volume. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

Clone 8WG16 recognizes YSPTSPS in the C-terminal domain. The antibody cross-reacts with wheat, yeast, mouse, c. elegans, x. laevis, and most eukaryotic RNAPII.

Additional reported applications (for the relevant formats) include: chromatin immunoprecipitation, functional assay, and immunoprecipatation.

Application References

(PubMed link indicates BioLegend citation)
  1. Edwards AM, et al. 1990. Proc. Natl. Acad. Sci. USA. 87:2122.
  2. Thompson NE, et al. 1990. J. Biol. Chem. 265:7069.
  3. Thompson NE, et al. 1989. J. Biol. Chem. 264:11511.
Product Citations
  1. Sump B, et al. 2022. Elife. 11: . PubMed
  2. Wang C, et al. 2023. Nat Commun. 14:1598. PubMed
  3. Onishi R, et al. 2020. Sci Adv. 6:00. PubMed
  4. Niu L, et al. 2021. Nat Genet. 53:1075. PubMed
  5. Edwards SL, et al. 2021. Cell Reports. 36(9):109623. PubMed
  6. Catala M, et al. 2019. Commun Biol. 2:211. PubMed
  7. Wang Y, et al. 2020. J Biol Chem. . PubMed
  8. Zhao X, et al. 2022. Nat Commun. 13:5401. PubMed
  9. Huang Z, et al. 2021. Molecular Cell. 81(5):953-968.e9. PubMed
  10. Di Giammartino DC, et al. 2022. Methods Mol Biol. 2532:113. PubMed
  11. Legrand N et al. 2019. Nucleic Acids Res. 47(18):9573-9591 . PubMed
  12. Montinaro A, et al. 2021. Cell Death Differ. Online ahead of print. PubMed
  13. Das S, et al. 2020. eLife. 9:00. PubMed
  14. Rowley MJ, et al. 2019. Cell Rep. 26:2890. PubMed
  15. Cardoso da Silva R, et al. 2020. PLoS Genet. 16:e1008905. PubMed
  16. Paakinaho V, et al. 2021. Nucleic Acids Res. 49:1951. PubMed
  17. Lee J et al. 2018. Cell stem cell. 22(3):428-444 . PubMed
  18. Vadivel CK, et al. 2021. Cancers (Basel). 13:00. PubMed
  19. Joo Y, et al. 2020. Nat Commun. 2.640972222. PubMed
  20. Rhie SK, et al. 2018. Genome Res. 13:e0196698. PubMed
  21. Thouvenot P, et al. 2016. J Cell Sci. 129(23):4366-4378. PubMed
  22. Fan X, et al. 2021. PLoS Genet. 17:e1009773. PubMed
  23. Yu Y, et al. 2016. Sci Rep. 6:31752. PubMed
  24. See YX, et al. 2022. Genome Res. . PubMed
RRID
AB_2565554 (BioLegend Cat. No. 664906)

Antigen Details

Structure
RPB1 is a 220 kD subunit of RNA polymerase II and has 1970 amino acids.
Distribution

Nucleus; ubiquitous expressed in all cells

Function
Core element of RNA polymerase II transcription machinery.
Interaction
COMPASS (COMplex of Proteins ASsociated with Set1), SET2, BRE1
Biology Area
Cell Biology, Transcription Factors
Molecular Family
Nuclear Markers
Antigen References
  1. Chávez S, et al. 2016. CUrr. Genet. 62:701.
Gene ID
5430 View all products for this Gene ID 20020 View all products for this Gene ID 2540292 View all products for this Gene ID
UniProt
View information about RNA Polymerase II RPB1 on UniProt.org

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Go To Top Version: 8    Revision Date: 07.09.2024

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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