Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody (bs-6581R)

Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) antibody

DATASHEET

Host:Rabbit

Target Protein:RNA polymerase II CTD repeat YSPTSPS (phospho S2)

IR:Immunogen Range:Y(p-S)PT

Clonality:Polyclonal

Isotype:IgG

Entrez Gene:5430

Swiss Prot:P24928

Source:KLH conjugated Synthesised phosphopeptide derived from human RNA polymerase II RPB1 around S2:Y(p-S)PT 

Purification:affinity purified by Protein A

Storage:0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol. Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.

Background: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. It is composed of mobile elements that move relative 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 stranded DNA template strand of the promoter is positioned within the central active site cleft of Pol II. 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. During transcription elongation, Pol II moves on the template as the transcript elongates. 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. Acts as a RNA-dependent RNA polymerase when associated with small delta antigen of Hepatitis delta virus, acting both as a replicate and transcriptase for the viral RNA circular genome.

Size:100ul

Concentration:1mg/ml

Applications:WB(1:500-2000)
ELISA(1:5000-10000)
IHC-P(1:100-500)
IHC-F(1:100-500)
Flow-Cyt(1ug/Test)
IF(1:100-500)

Cross Reactive Species:Human
Mouse
Rat
Dog
Cow
.

For research use only. Not intended for diagnostic or therapeutic use.

VALIDATION IMAGES

Tissue/cell: rat colon tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-RNA polymerase II CTD repeat YSPTSPS (phospho S2) Polyclonal Antibody, Unconjugated(bs-6581R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining

Blank control: HL60. Primary Antibody (green line): Rabbit Anti-RNA polymerase II CTD repeat YSPTSPS antibody (bs-6581R-FITC) Dilution: 0.2μg /10^6 cells; Isotype Control Antibody (orange line): Rabbit IgG-FITC . Protocol The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at-20℃. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at at room temperature . The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.

Blank control: HL60.
Primary Antibody (green line): Rabbit Anti-RNA polymerase II CTD repeat YSPTSPS antibody (bs-6581R-FITC)
Dilution: 0.2μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG-FITC .
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at-20℃. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at at room temperature . The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.