Cat. No.
MABL-2840
Application
CHIP, ChIP-seq, FACS, IP, WB, ELISA, IF, IHC
Isotype
Engineer antibody
Species Reactivity
Saccharomyces cerevisiae, Human
Clone No.
CTD 4H8
From
Recombinant Antibody
Specificity
CTD 4H8 is known to be reactive with human and S. cerevisiae RNA polymerase II CTDs, however may also be cross-reactive with this protein in other species, including mouse, rat and chicken. The Ab is also reactive with both the phosphorylated and unphosphorylated form of the CTD YSPTSPS repeat. RNAPII is responsible for the transcription of most protein-coding genes in the human genome. The CTD of the enzyme, featuring these YSPTSPS repeats, is crucial for the regulation of the enzyme in a spatial (i.e. positionally along genes) and temporal (i.e. rate of RNA polymerisation) manner.
Alternative Names
RNAPII; DNA-directed RNA polymerase subunit RPB1; RNA polymerase II subunit B1
UniProt
P24928
Immunogen
Murine antibodies were produced by immunising mice with a peptide consisting of 10 repeats of the RNA polymerase II C-terminal domain sequence YSPTSPS. In each repeat, Ser5 was phosphorylated. Immunised cells were fused with the Sp2/0-Ag14 myeloma cell line to produce a hybridoma.
Application Notes
This mAb has been used in a variety of methods, including Western blotting of the unphosphorylated (Engelhardt et al, 2005) or Ser5-CTD phosphorylated form (Stock et al, 2007), and in precipitation experiments either simply pulling down RNAPII (Kukalev et al, 2005; Peng et al, 2014) or coupled to massively parallel DNA sequencing (ChIP-seq) of the total RNAPII-associated DNA isolated from human CD4+ cells (Zhang et al, 2012).
Antibody First Published
Kristjuhan et al. Transcriptional inhibition of genes with severe histone h3 hypoacetylation in the coding region Mol Cell. 2002 Oct;10(4):925-33 PMID:12419235
Note on publication
Describes a growing link between histone acetylation and gene expression using budding yeast. The mAb (CTD) 4H8 was used to immunoprecipitate hypo- and hyperphosphorylated RNAPII. The reactivity of CTD 4H8 for these forms of RNAPII was described by B. Winkler and J.Q.S, but the experiments which determined this, or the specific production of this mAb, remain unpublished.
Size
100 μg Purified antibody.
Concentration
1 mg/ml.
Purification
Protein A affinity purified
Buffer
PBS with 0.02% Proclin 300.
Storage Recommendation
Store at 4⁰C for up to 3 months. For longer storage, aliquot and store at - 20⁰C.
