Rat Phospho-POLR2A (Ser5) Ready-To-Use IHC Kit

Cat. No.: IHC0626R
Sample Type: FFPE tissue
Size: 50T (including a control slide)
Storage and Stability: Please store components at the temperatures indicated on the individual tube labels. The kit is stable for 6 months from the date of receipt.

Background

DNA-directed RNA polymerase II subunit RPB1 (POLR1A) is a DNA-dependent RNA polymerase that catalyzes the transcription of DNA into RNA using the four ribonucleoside triphosphates as substrates. POLR1A is the largest subunit and is a catalytic component of RNA polymerase II which synthesizes mRNA precurors and many functional non-coding RNAs. It also 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 though 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 stie and is through 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's largest subunit (RPB1), which serves as a platform for assembly of factors that regulate transcription initiation, elongation, termination, and mRNA processing. Regulation of gene expression levels depends on the balance between methylation and acetylation levelts of the CTD-lysines.

Synonyms

220kDa; hRPB220; hsRPB1; POLR2; POLR2A; POLRA; RPB1; RPBh1; Rpii215; RpIILS; RPO2; Rpo2-1; RPOL2.

Materials Supplied

Number	Component	50T	Concentration	Storage
1	PBS Buffer (powder)	2L x 2	20x	RT
2	Antigen Retrieval Buffer	20ml	100x	2-8°C
3	Endogenous Peroxidase Blocking Buffer	3ml	RTU	2-8°C
4	Blocking Buffer	3ml	RTU	2-8°C
5	Primary Antibody (Rat Phospho-POLR2A (Ser5) Recombinant Rabbit mAb)	6ml	RTU	2-8°C
6	Secondary Antibody (Goat Anti-Rabbit IgG H&L, HRP conjugated)	6ml	RTU	2-8°C
7	Chromogen Component A	0.3ml	RTU	-20°C
8	Chromogen Component B	0.3ml	RTU	-20°C
9	Counter Staining Reagent	5ml	RTU	RT
10	Differentiation Reagent	6ml	RTU	RT
11	Mounting Media	5ml	RTU	RT
12	Control slide (Rat colon)	1 slide	RTU	RT
13	Datasheet	1 copy

Notes

1. The positive control slide provided in the kit allows you to be sure that the experimental set-up is working properly.
2. Do not allow slides to dry at any time during this procedure.
3. Please don't replace the matching reagents in this product with other manufacturers' products.
4. As DAB is a carcinogen, please take necessary precautions.
5. PBS (reagent 1) can be stored for one week at 4℃ after preparation; The antigen retrieval buffer (1×reagent 2) and the chromogenic agent (the mixture of reagents 7 and 8) should be prepared right before each assay.

As DAB is a carcinogen, please take necessary precautions.

Important Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.