Polymerase (RNA) II (DNA Directed) Polypeptide A, 220kDa (POLR2A) (pSer5) antibody

Details for Product No. ABIN112080
Request Want additional data for this product?

The Independent Validation Initiative strives to provide you with high quality data. Find out more

Antigen
Synonyms NCU02103.1, 19.m02399, POLR2A, 220kDa, Rpb1, Rpo2-1, POLR2, POLRA, RPB1, RPBh1, RPO2, RPOL2, RpIILS, hRPB220, hsRPB1
Epitope
pSer5
(11), (11), (10), (5), (3), (3), (2), (2), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1), (1)
Reactivity
Human, Mouse (Murine), Yeast
(57), (21), (21), (2), (1), (1), (1), (1)
Host
Mouse
(42), (18), (1), (1)
Clonality (Clone)
Monoclonal ()
Application
ELISA, Immunocytochemistry (ICC), Immunofluorescence (IF), Immunoprecipitation (IP), Western Blotting (WB)
(56), (30), (27), (22), (13), (8), (5), (4), (3), (3), (1), (1), (1)
Pubmed 4 references available
Quantity 50 μL
Shipping to United States (Change)
Availability Will be delivered in 6 to 8 Business Days
Request Want additional data for this product?

The Independent Validation Initiative strives to provide you with high quality data. Find out more

Catalog No. ABIN112080
544.50 $
Plus shipping costs $45.00

Order hotline:

  • +1 404 474 4654
  • +1 888 205 9894 (TF)
Immunogen 10 repeats of synthetic peptide: YSPTSPS using chemically synthesized phosphor-Ser5.
Clone 4H8
Isotype IgG1
Specificity Recognises the C-terminal repeat of the largest subunit of RNA polymerase II. It recognises both unphosphorylated and phosphorylated forms. Cross-reacts with Human and S. cerevisiae. Expected to cross-react with Drosophilia melanogaster, Hamster, Mouse, S. pombe, Arabidopsis thaliana due to sequence homology - all of these species contain at least one or more 100% identical repeats.
Purification Protein A affinity
Alternative Name POLR2A
Background RNA polymerase II carboxy-terminal domain (CTD) interacts with a large multisubunit complex that contains TATA-binding protein (TBP) and is an integral part of the transcription initiation complex. Phosphorylation of RNA polymerase II's largest subunit C- terminal domain (CTD) is a key event during mRNA metabolism. Numerous enzymes, including cell cycle-dependent kinases and TFIIF-dependent phosphatases target the CTD.
Alternate names: DNA-directed RNA polymerase II largest subunit, DNA-directed RNA polymerase II subunit RPB1, POLR2, POLRA, RNA polymerase II, RNA polymerase II 220 kd, RNA polymerase II subunit B1, RNA-directed RNA polymerase II subunit RPB1, RPB1, RPB220, RPOL2
Gene ID 5430
NCBI Accession NP_000928
UniProt P24928
Application Notes Suitable for Western blot, Immunoprecipitation, ELISA and Immunofluorescence. Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.
Protocol Immunofluorescence protocol-Formaldehyde fixationCollect cells from T. c. unit and remove media from petri dish using suction. Wash with 1x PBS and remove. Incubate cells in pre-warm (37°C) Para-Formaldehyde for 12 minutes at room temperatureon an orbital shaker. Remove PFA and incubate in 0. 5% Triton X-IOO in 1x PBS for 5 minutes at roomtemperature. Prepare blocking reagent, this is also the antibody diluent. Wash cells 2x with 1x PBS at room temperature, for 4 minutes/wash on an orbital shaker. Block with 1 % NCS and 1x PBS for 30 minutes at room temperature. Prepare primary antibodies (50μl/coverslip) and moist staining chambers. Wash cells 2x with lx PBS at room temperature and air dry briefly. Incubate with primary antibody for 1 hr at room temperature in the dark in stainingchambers. During this time prepare the secondary antibody. Wash cells 5x with 1x PBS (5 beaker changes/5 counts in each beaker)Incubate with secondary antibody for 1 hour at room temperature in the dark in stainingchambers. Wash cells 5x with 1x PBS. Mount in Dapi. Solutions (prepare fresh the same day of staining). 1x Phosphate buffered saline. Blocking reagent: 1% NCS in 1x PBS (use fresh l0x PBS). Fixation solution: 3. 5% Para formaldehyde. 1. 75g PFA in 20 ml d. H20 plus 5 drops 1M NaOH. Stir on a hot plate at 50-60°C untildissolved. Add 4 drops IN HCI and check pH indicator strip. PH should be 7. 4. Completevolume with d. H20 to 25ml and add 25ml 2xPBS. Check pH before adding to cover slips. Immunofluorescence protocol - Methanol/acetone fixationCollect cells from T. C. unit and remove media from petri dish using suction. Wash with 1x PBS and remove. Fix cells with cold methanol: acetone 1: 1 for 10 minutes on ice. Prepare blocking reagent, this is also the diluent for the antibodies. Remove fixative and wash cells 3x with Ix PBS at RT, for 4 minutes/wash on orbital shaker. Block with 1% NCS and Ix PBS for 30 minutes at RT. Prepare primary antibodies (50μl/coverslip) and moist staining chambers. Wash cells 2x with 1 x PBS at RT and air dry for approximately 7 minutes. Incubate with primary antibody for 1 hr at RT in the dark in staining chambers. During thistime prepare secondary antibody. Wash cells 5x with 1x PBS (5 beaker changes/5 counts in each beaker)Incubate with secondary antibody for 1 hr at R T in the dark in staining chambers. Wash cells 5x with 1x PBS. Mount in Dapi. Solutions (prepare fresh the same day of staining)1x Phosphate buffered saline. Blocking reagent: 1% NCS in 1x PBS (use fresh 10x PBS). Fixation solution: methanol: acetone 1: 1 ice cold.
Restrictions For Research Use only
Format Liquid
Concentration 0.8 mg/mL
Buffer PBS pH 7.2 with 0.09% sodium azide as preservative.
Preservative Sodium azide
Precaution of Use This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handling Advice Avoid repeated freezing and thawing.
Storage -20 °C
Expiry Date 12 months
Background publications Tomaschewski, Gram, Crabb et al.: "T4-induced alpha- and beta-glucosyltransferase: cloning of the genes and a comparison of their products based on sequencing data." in: Nucleic acids research, Vol. 13, Issue 21, pp. 7551-68, 1985 (PubMed).

Patturajan, Conrad, Bregman et al.: "Yeast carboxyl-terminal domain kinase I positively and negatively regulates RNA polymerase II carboxyl-terminal domain phosphorylation." in: The Journal of biological chemistry, Vol. 274, Issue 39, pp. 27823-8, 1999 (PubMed).

Kristjuhan, Walker, Suka et al.: "Transcriptional inhibition of genes with severe histone h3 hypoacetylation in the coding region." in: Molecular cell, Vol. 10, Issue 4, pp. 925-33, 2002 (PubMed).

Kristjuhan, Wittschieben, Walker et al.: "Spreading of Sir3 protein in cells with severe histone H3 hypoacetylation." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 100, Issue 13, pp. 7551-6, 2003 (PubMed).

Validation Images
Did you look for something else?
back to top