Product Name:
IWS1-S720+T721+T725
Product Number:
ab-pn611
Target Full Name: Protein IWS1 homologue
Target Alias: DKFZp761G0123; IWS1L; Protein IWS1 homolog: IWS1-like protein
Product Type Specific: IWS1 phosphosite-specific antibody
Antibody Code: PN611
Antibody Target Type: Phosphosite-specific
Antibody Phosphosite: S720+T721+T725
Protein UniProt: Q96ST2
Protein SigNET: IWS1
Antibody Type: Polyclonal
Antibody Host Species: Rabbit
Antibody Immunogen Source: Synthetic phosphopeptide patterned after human IWS1
Antibody Immunogen Sequence: RMN(pS)(pT)GGQ(pT)PRRC
Antibody Immunogen Description: Corresponds to amino acid residues R717 to R728. S720 and T721 phosphorylation inhibit interactions with SPT6 and THOC4, and promote cell motility. The effects of T725 phosphorylation are unclear. These are three of the major in vivo phosphorylation sites in ITSN2 (≥804, ≥83 and ≥253 reports, respectively, from high throughput mass spectrometry studies recorded in PhosphoSitePlus). IWS1 is known to be phosphorylated in vitro at S720 and T721 by Akt1, and Akt3.
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.
Antibody Modification: Unconjugated. Contact KInexus if you are interest in having the antibody biotinylated or coupled with fluorescent dyes.
Antibody Concentration: 0.75 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 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 | Platypus | Chicken | Frog.
Scientific Background: IWS1 (Interacts with Spt6 1) is a transcription accesroy protein that belongs in to the IWS1 family. It is an essential RNA polymerase II (Pol II)-associated transcription elongation factor that acts as a nuclear, scaffolding protein to couple transcription with pre-mRNA processing, splicing, and export. It is recruited to the RNA polymerase II (RNAPII) elongation complex via binding to the transcription elongation factor SUPT6H and the C-terminal domain (CTD) of the RNAPII subunit RPB1 (POLR2A). The SUPT6H:IWS1:CTD complex recruits mRNA export factors (ALYREF/THOC4, EXOSC10) as well as histone modifying enzymes (such as SETD2) to ensure proper mRNA splicing, efficient mRNA export and elongation-coupled H3K36 methylation, a signature chromatin mark of active transcription. It mediates H3K36 trimethylation by recruiting the histone methyltransferase SETD2. It features a central HEAT/TND domain that interacts with the elongation complex, and flanked by disordered N- and C-terminal regions. IWS1 may function as a scaffold that positions downstream DNA within the cleft of Pol II to stimulate elongation velocity. IWS1 activity is modulated by phosphorylation by Akt isoforms, which affects alternative splicing of target genes. It is crucial for cell survival and development, as its loss causes pre-implantation lethality. This description may include information annotated by UniProt and/or Google AI.