Key features and details | |
Cat. No. | MABL-2840 |
Name | Anti-RNA polymerase II mAbs |
Clone No. | AFD- CTD 4H8 |
From | Recombinant Antibody |
Isotype | Engineer antibody |
Application | CHIP, ChIP-seq, FACS, IP, WB, ELISA, IF, IHC |
Species Reactivity | Saccharomyces cerevisiae, Human |
Basic Information | |
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 Name | 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by B. Winkler and J.Q.S, but the experiments which determined this, or the specific production of this mAb, remain unpublished. |
COA Information （For reference only, actual COA shall prevail） | |
Size | 100 μg Purified antibody. |
Concentration | 1 mg/ml. |
Purification | Protein A affinity purified |
Buffer | PBS with 0.02% Proclin 300. |
Concentration | 1 mg/ml. |
Storage Recommendation | Store at 4⁰C for up to 3 months. For longer storage, aliquot and store at - 20⁰C. |
