Product Details

Full Product Name

Rabbit anti-Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (Baker's yeast) RPO21 Polyclonal antibody

Uniprot No.

P04050

Target Names

RPO21

Alternative Names

RPO21 antibody; RPB1 antibody; RPB220 antibody; SUA8 antibody; YDL140C antibody; D2150 antibody; DNA-directed RNA polymerase II subunit RPB1 antibody; RNA polymerase II subunit 1 antibody; RNA polymerase II subunit B1 antibody; EC 2.7.7.6 antibody; DNA-directed RNA polymerase III largest subunit antibody; RNA polymerase II subunit B220 antibody

Raised in

Rabbit

Species Reactivity

Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (Baker's yeast)

Immunogen

Recombinant Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (Baker's yeast) RPO21 protein

Immunogen Species

Saccharomyces cerevisiae (strain ATCC 204508 / S288c) (Baker's yeast)

Conjugate

Non-conjugated

Clonality

Polyclonal

Isotype

IgG

Purification Method

Antigen Affinity Purified

Concentration

It differs from different batches. Please contact us to confirm it.

Buffer

Preservative: 0.03% Proclin 300
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4

Form

Liquid

Tested Applications

ELISA, WB (ensure identification of antigen)

Protocols

ELISA Protocol

Troubleshooting and FAQs

Antibody FAQs

Storage

Upon receipt, store at -20°C or -80°C. Avoid repeated freeze.

Value-added Deliverables

① 200ug * antigen (positive control);
② 1ml * Pre-immune serum (negative control);

Quality Guarantee

① Antibody purity can be guaranteed above 90% by SDS-PAGE detection;
② ELISA titer can be guaranteed 1: 64,000;
③ WB validation with antigen can be guaranteed positive;

Lead Time

Made-to-order (14-16 weeks)

Usage

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

Target Background

Function

DNA-dependent RNA polymerase catalyzes the transcription of DNA into RNA using the four ribonucleoside triphosphates as substrates. Largest and catalytic component of RNA polymerase II which synthesizes mRNA precursors and many functional non-coding RNAs. Forms the polymerase active center together with the second largest subunit. Pol II is the central component of the basal RNA polymerase II transcription machinery. During a transcription cycle, Pol II, general transcription factors and the Mediator complex assemble as the preinitiation complex (PIC) at the promoter. 11-15 base pairs of DNA surrounding the transcription start site are melted and the single-stranded DNA template strand of the promoter is positioned deeply within the central active site cleft of Pol II to form the open complex. After synthesis of about 30 bases of RNA, Pol II releases its contacts with the core promoter and the rest of the transcription machinery (promoter clearance) and enters the stage of transcription elongation in which it moves on the template as the transcript elongates. Pol II appears to oscillate between inactive and active conformations at each step of nucleotide addition. Elongation is influenced by the phosphorylation status of the C-terminal domain (CTD) of Pol II largest subunit (RPB1), which serves as a platform for assembly of factors that regulate transcription initiation, elongation, termination and mRNA processing. Pol II is composed of mobile elements that move relative to each other. The core element with the central large cleft comprises RPB3, RBP10, RPB11, RPB12 and regions of RPB1 and RPB2 forming the active center. The clamp element (portions of RPB1, RPB2 and RPB3) is connected to the core through a set of flexible switches and moves to open and close the cleft. A bridging helix emanates from RPB1 and crosses the cleft near the catalytic site and is thought to promote translocation of Pol II by acting as a ratchet that moves the RNA-DNA hybrid through the active site by switching from straight to bent conformations at each step of nucleotide addition. In elongating Pol II, the lid loop (RPB1) appears to act as a wedge to drive apart the DNA and RNA strands at the upstream end of the transcription bubble and guide the RNA strand toward the RNA exit groove located near the base of the largely unstructured CTD domain of RPB1. The rudder loop (RPB1) interacts with single-stranded DNA after separation from the RNA strand, likely preventing reassociation with the exiting RNA. The cleft is surrounded by jaws: an upper jaw formed by portions of RBP1, RPB2 and RPB9, and a lower jaw, formed by RPB5 and portions of RBP1. The jaws are thought to grab the incoming DNA template, mainly by RPB5 direct contacts to DNA.

Gene References into Functions

Here, the authors identify a site of ubiquitination (K1246) in the catalytic subunit of RNAPII close to the DNA entry path. Ubiquitination was increased in the absence of the Bre5-Ubp3 ubiquitin protease complex. PMID: 29027900
The authors determined that the tandem SH2 domain of Saccharomyces cerevisiae Spt6 binds the linker region of the RNA polymerase II subunit Rpb1 rather than the expected sites in its heptad repeat domain. The 4 nM binding affinity requires phosphorylation at Rpb1 S1493 and either T1471 or Y1473. PMID: 28826505
Data suggest the binding motif in the RPB1 (RPO21) switch 1 of RNA polymerase II (pol II) can sense the structural changes of the DNA minor groove (minor groove sensor) before duplex unwinding. PMID: 27791148
RPO21 interaction with LYS2 and Spt elongation factors PMID: 27261007
Promoter and template-specific effects on severity of gene expression defects have been identified for both fast and slow Pol II mutants. PMID: 28119420
Studies indicate that the covalent, flexible linker ensured that transcription factor TFIIB (TFIIB) was recruited stoichiometrically near its binding site on the RNA polymerase (Pol) II (Pol II) surface. PMID: 27528766
RPB1 mutants in the foot region of RNA polymerase II affect the assembly of the complex by altering the correct association of both the Rpb6 and the Rpb4/7 dimer PMID: 27001033
Sen1, Nrd1 and Ysh1 play a role in RNA polymerase II termination. PMID: 25299594
Results indicate that multiple mechanistic features contribute to 5'-3' exoribonuclease Rat1-mediated termination of RNA polymerase II (RNAPII). PMID: 25722373
The results show that the Rpb4/7 heterodimer in Saccharomyces cerevisiae plays a key role in controlling phosphorylation of the carboxy terminal domain of the Rpb1 subunit of RNA polymerase II. PMID: 25416796
Data indicate reduced promoter association of Mediator and TATA-binding protein in a RNA polymerase II (rpb1-1) mutant. PMID: 24727477
These results show that, with some spatial constraints, CTD can function even when disconnected from Rpb1. PMID: 24035501
Results indicate an important role of the intrinsic blocks to forward translocation in pausing by RNA polymerase II (RNAP II). PMID: 23238253
RPB1 (RPO21) mutations increase transcriptional slippage in S. cerevisiae PMID: 23223234
Data show that polymerases transcribing opposite DNA strands cannot bypass each other, and head-to-head collision results in RNA polymerase II (RNAPII) stopping. PMID: 23041286
find that the transcription processivity defect imposed on RNAPII by the rpb1-N488D mutation is corrected upon Rat1p inactivation. PMID: 20188675
RPB1 has a role in hypersensitivity to 6-azauracil PMID: 16510790
An Rsp5 dimer assembled on the RNA polymerase II (RNAP II) C-terminal domain; ubiquitylation sites bind Ubc5, which in turn binds Rsp5 to allow modification of RPB1 subunit of RNAP II. PMID: 17418786
In response to ultraviolet radiation, Rpb9 also functions in promoting ubiquitylation and degradation of Rpb1, the largest subunit of Pol II. PMID: 17452455
Saccharomyces cerevisiae RNA polymerase II (Pol II) "trigger loop" participates in substrate selection PMID: 18538653
mutant showed a 10-fold decrease in fidelity of transcription elongation in vitro PMID: 18538654
Rpb1p interacts with a large number of proteins involved in mRNA synthesis, processing, export, and other cellular processes. PMID: 19193966
results demonstrate a novel covalent modification of Rpb1 in response to UV induced DNA damage or transcriptional impairment, and unravel an important link between the modification and the DNA damage checkpoint response PMID: 19384408
Data show that Rpb1 CTD serine 7 is also phosphorylated and that Ser-7(P) chromatin immunoprecipitation patterns resemble those of Ser-5(P). PMID: 19679665

Hide All

Subcellular Location

Nucleus.

Protein Families

RNA polymerase beta' chain family

Database Links

KEGG: sce:YDL140C

STRING: 4932.YDL140C

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RPO21 Antibody

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