Transcription Factor IIF, Rap74 subunit

• Species: Human
• Expression Host: E. Coli
• Tag: His-tag
• Purity: 90%
• Molecular Weight: 59.3 kDa.
• Gene Accession Number: NM_002096.

Applications

1 unit equals 1 nanogram of purified protein. 1 - 10 units are sufficient for a gel mobility shift assay in a 20 µl reaction; 20 units are sufficient for a reconstituted transcription assay and 100 units are sufficient for a protein-protein interaction assay. For research use only.

Formulation and Storage

Protein is stored in 20 mM Tris-Cl (pH 8.0), 20% Glycerol, 100 mM KCl, 1 mM DTT and 0.2 mM EDTA buffer and kept at -800C. Avoid repeated freeze thaw cycles.

Target Gene Sequence

MAALGPSSQNVTEYVVRVPKNTTKKYNIMAFNAADKVNFATWNQ ARLERDLSNKKIYQEEEMPESGAGSEFNRKLREEARRKKYGIVLKEFRPEDQPWLLRV NGKSGRKFKGIKKGGVTENTSYYIFTQCPDGAFEAFPVHNWYNFTPLARHRTLTAEEA EEEWERRNKVLNHFSIMQQRRLKDQDQDEDEEEKEKRGRRKASELRIHDLEDDLEMSS DASDASGEEGGRVPKAKKKAPLAKGGRKKKKKKGSDDEAFEDSDDGDFEGQEVDYMSD GSSSSQEEPESKAKAPQQEEGPKGVDEQSDSSEESEEEKPPEEDKEEEEEKKAPTPQE KKRRKDSSEESDSSEESDIDSEASSAFFMAKKKTPPKRERKPSGGSSRGNSRPGTPSA EGGSTSSTLRAAASKLEQGKRVSEMPAAKRLRLDTGPQSLSGKSTPQPPSGKTTPNSG DVQVTEDAVRRYLTRKPMTTKDLLKKFQTKKTGLSSEQTVNVLAQILKRLNPERKMIN DKMHFSLKE

SYNONYM

BTF4; RAP74; TF2F1; TFIIF; Homo sapiens general transcription factor IIF, polypeptide 1, 74kDa (GTF2F1).

Background

The transcription factor IIF (TFIIF) is a heterodimer, composed of 58 kDa (RAP74) and 26 kDa (RAP30) subunits. It was first identified through the ability to interact with immobilized RNA polymerase II (1, 2). In addition to its role in transcription initiation, TFIIF can increase the specificity and efficiency of RNA polymerase II transcription, and can especially increase the rate of transcription elongation (3, 4).
The His tagged recombinant RAP74 protein is isolated from an E. coli strain that carries the coding sequence of human RAP74 under the control of a T7 promoter.
RAP74 is required for stimulation of the rate of RNA
polymerase II elongation through a direct interaction with pol II and for start site selection. RAP74 is heavily phosphorylated in vivo and can be phosphorylated by TAFII250, a subunit of TFIID (5).

Molecular weight is calculated by protein sequence of full length Rap74. SDS_PAGE migration is around 70 kDa.

SDS-PAGE with Coomassie Blue Staining