Product Name:
GTF2F1-pS385+pT389
Product Number:
ab-pk651
Target Full Name: General transcription factor IIF subunit 1
Target Alias: BTF4; General transcription factor IIF polypeptide 1 74 kDa subunit protein; General transcription factor IIF, polypeptide 1 (74kD subunit); General transcription factor IIF, polypeptide 1, 74kDa; RAP74; T2FA; TF2F1; TFIIF; Transcription initiation factor IIF alpha subunit; Transcription initiation factor IIF subunit alpha; Transcription nitiation factor RAP74
Product Type Specific: GTF2F1 protein kinase phosphosite-specific antibody
Antibody Code: PK651
Antibody Target Type: Phosphosite-specific
Antibody Phosphosite: S385+T389
Protein UniProt: P35269
Protein SigNET: GTF2F1
Antibody Type: Polyclonal
Antibody Host Species: Rabbit
Antibody Immunogen Source: Human GTF2F1 sequence peptide Cat. No.: PE-04AMS95
Antibody Immunogen Sequence: RGN(pS)RPG(pT)PSA(bA)C
Antibody Immunogen Description: Corresponds to amino acid residues R382 to A392; in the TFIIF_alpha domain. S385+T389 phosphorylation inhibits transcriptional activity. These are the two major in vivo phosphorylation sites in GTF2F1 (≥280 and ≥319 reports, respectively, from high throughput mass spectrometry studies recorded in PhosphoSitePlus). GTF2F1 is known to be phosphorylated in vitro at S385 by GTF2F1, and at T389 by GTF2F1.
Production Method: The immunizing peptide was produced by solid phase synthesis on a multipep peptide synthesizer and purified by reverse-phase hplc chromatography. Purity was assessed by analytical hplc and the amino acid sequence confirmed by mass spectrometry analysis. This peptide was coupled to KLH prior to immunization into rabbits. New Zealand White rabbits were subcutaneously injected with KLH-coupled immunizing peptide every 4 weeks for 4 months. The sera from these animals was applied onto an agarose column to which the immunogen peptide was thio-linked. Antibody was eluted from the column with 0.1 M glycine, pH 2.5. Subsequently, the antibody solution was neutralized to pH 7.0 with saturated Tris.This antibody was also subject to negative purification over phosphotyrosine-agarose.
Antibody Modification: Unconjugated. Contact KInexus if you are interest in having the antibody biotinylated or coupled with fluorescent dyes.
Antibody Concentration: 1 mg/ml
Storage Buffer: Phosphate buffered saline pH 7.4, 0.05% Thimerasol
Storage Conditions: For long term storage, keep frozen at -40°C or lower. Stock solution can be kept at +4°C for more than 3 months. Avoid repeated freeze-thaw cycles.
Product Use: Western blotting | Antibody microarray
Antibody Dilution Recommended: 2 µg/ml for immunoblotting
Antibody Potency: Medium immunoreactivity of a target-sized protein by Western blotting in maturing, germinal vesicle positive, sea star oocytes. Very strong immunoreactivity with immunogen peptide on dot blots.
Antibody Species Reactivity: This antibody detects the target phosphoprotein in the following species due to conservation of amino acid sequence: Human | Chimpanzee | Rhesus macaque | Dog | Rat | Mouse | Frog.
Antibody Positive Control: The observed molecular mass of the processed target protein on SDS-PAGE gels is reported to be around 66-74 kDa.
Antibody Specificity: High-very high
Antibody Cross Reactivity: No significant cross-reactive proteins detected in A431, Jurkat and MCF7 cells, except for a strong immunoreactive ~75 kDa protein in FBS-treated A431 cells and germinal vesicle positive-maturing sea star oocytes.
Related Product 1: GTF2F1 (382-392) pS385+pT389 antibody blocking peptide (Cat. No.: PE-04AMS95)
Related Product 2: GTF2I S412 phosphorylation antibody (Cat. No.: AB-PN602)
Scientific Background: GTF2F1 (General Transcription Factor IIF Subunit 1) is a 74 kDa protein (RAP74) that acts as the alpha subunit of TFIIF, a crucial component of the RNA polymerase II transcription machinery. As part of the TFIIF complex with the smaller 30 kDa GTF2F2 subunit, GTF2F1 facilitates the binding of RNA polymerase II to promoter regions, working in conjunction with TFIIB after TFIID has bound to the DNA It also promotes transcriptional elongation. GTF2F1 interacts with other several factors, including POLR2A, TATA-binding protein, the N-terminal region of RAP30, TAFII250, and the androgen receptor. It is also a protein-serine/threonine kinase of the Atypical group and family. It can be inhibited by mutation of Ser-385 and Thr-389, which are autophosphorylation sites. It is also identified as a substrate of TAFII250. GTF2F1 also plays a role in the cellular response to viruses. This description may include information annotated by UniProt and/or Google AI.