Product Name:
WT1-pS365
Product Number:
ab-pn732
Target Full Name: Wilms tumour protein
Target Alias: WT33
Product Type Specific: WT1 phosphosite-specific antibody
Antibody Code: PN732
Antibody Target Type: Phosphosite-specific
Antibody Phosphosite: S365
Protein UniProt: P19544
Protein SigNET: WT1
Antibody Type: Polyclonal
Antibody Host Species: Rabbit
Antibody Immunogen Source: Synthetic phosphopeptide patterned after human WT1
Antibody Immunogen Sequence: CERRF(pS)RSD
Antibody Immunogen Description: Corresponds to amino acid residues E361 to D368. The effect of S365 phosphorylation is unclear. In vivo phosphorylation of WT1 has not been reported for this phosphosite (0 reports from high throughput mass spectrometry studies recorded in PhosphoSitePlus). This human phosphosite is highly conserved in vertebrates and also found in fish. A nearly identical amino acid sequence surrounds S393. WT1 is known to be phosphorylated at this site in vitro by PKACa (PRKACA).
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: 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 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 | Chicken | Zebra fish.
Scientific Background: WT1 (Wilms tumour protein) is a multifunctional zinc-finger transcription factor that belongs to the EGR C2H2-type zinc-finger protein family. It plays an important role in development of kidneys, gonads, spleen, and heart, and cell survival, by acting as both a transcriptional activator and as a repressor. It recognizes and binds to the DNA promoter sequence 5'-GCG(T/G)GGGCG-3'. It regulates the expression of numerous target genes, including EPO. It serves an important role for development of the urogenital system. It has a tumour suppressor as well as an oncogenic role in tumour formation. Its function may be isoform-specific. It features four Kruppel-type C2-H2 zinc fingers at the C-terminus and is rich in proline, glutamine, and glycine residues. Alternative splicing creates four major isoforms, commonly defined by the presence or absence of a 17-amino acid segment (exon 5) and a 3-amino acid sequence (KTS). Isoforms lacking the KTS motif may act as transcription factors. Isoforms containing the KTS motif may bind mRNA and play a role in mRNA metabolism or splicing. Isoform 1 has lower affinity for DNA, and can bind RNA. It is expressed in the kidney and a subset of hematopoietic cells. WT1 acts as an oncogene in many cancers (e.g., leukemia, mesothelioma, breast cancer) by inhibiting apoptosis. Mutations lead to Wilms' tumour, Denys-Drash syndrome, and Frasier syndrome. This description may include information annotated by UniProt and/or Google AI.