Product Name:
TRIM28-pY517
Product Number:
ab-pk835
Target Full Name: Transcription intermediary factor 1-beta
Target Alias: KAP-1; KRIP-1; RING finger protein 96; RNF96; TIF1-beta; Tripartite motif-containing protein 28; TRIM28
Product Type Specific: TRIM28 protein kinase phosphosite-specific antibody
Antibody Code: PK835
Antibody Target Type: Phosphosite-specific
Antibody Phosphosite: Y517
Protein UniProt: Q13263
Protein SigNET: TRIM28
Antibody Type: Polyclonal
Antibody Host Species: Rabbit
Antibody Immunogen Source: Human TRIM28 (TIF1B) sequence peptide Cat. No.: PE-04APY99
Antibody Immunogen Sequence: TED(pY)NLI(bA)C
Antibody Immunogen Description: Corresponds to amino acid residues T514 to I520; in the region between the zf-B_box and PHD domains. The major site of phosphorylation in TRIM28.Y517 phosphorylation stimulates interaction with HP1 alpha and induces DNA repair. This is a major site of in vivo phosphorylation in TRIM28 based on ≥1375mass spectrometry report recorded in PhosphoSitePlus. TRIM28 is known to be phosphorylated at this site in vitro by Lyn.
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: 1.0 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 | Dog | Rat | Mouse.
Antibody Positive Control: The observed molecular mass of the processed target protein on SDS-PAGE gels is reported to be around 100-120 kDa.
Antibody Specificity: High-very high
Antibody Cross Reactivity: No significant cross-reactive proteins in A431, HeLa and MCF7 cells.
Related Product 1: TIF1b (514-520) pY517 antibody blocking peptide (Cat. No.: PE-04APY99)
Related Product 2: TRIM28-pY458 phosphosite-specific antibody (Cat. No.: AB-PK834)
Related Product 3: TRIM33 S1119 phosphorylation antibody (Cat. No.: AB-PK836)
Scientific Background: TRIM28 (Tripartite Motif-containing 28; TIF1b) is a ~110 kDa protein-serine/threonine kinase of the Atypical group and TIF1 family. It is a multifunctional scaffolding protein that acts as a corepressor, regulating transcription, chromatin remodeling, and DNA damage repair. It is essential for maintaining genome stability by silencing transposable elements. It features an N-terminal RBCC domain (RING, B-boxes, and Coiled-coil) that facilitates the formation of complexes, and a C-terminal PHD-bromodomain. It is recruited by KRAB domain-containing zinc finger proteins (KRAB-ZFPs) and acts as a universal co-repressor to establish heterochromatin, often by recruiting histone methyltransferase SETDB1. In response to DNA damage, it is phosphorylated by ATM to facilitate heterochromatin relaxation and repair of double-strand breaks. TRIM28 functions as a SUMO E3 ligase, regulating the stability of proteins like NLRP3 and contributing to stem cell pluripotency. It plays a crucial role in regulating the transcription of genes in activated CD8+ T cells. TIF family members are associated in retroviral restriction. TRIM28 contributes to EMT via E-cadherin and N-cadherin regulation in lung cancer cell lines. TRIM28 functions as both a tumor promoter and suppressor depending on the context, and it has been shown to regulate the growth and migration of tumour cells, such as in endometrial carcinoma and glioblastoma. This description may include information annotated by UniProt and/or Google AI.