Kinexus Products

Target Full Name: Heat shock 90 kDa protein beta

Target Alias: D6S182; FLJ26984; Heat shock 84 kDa; heat shock 90kD protein 1, beta; heat shock 90kDa protein 1, beta; heat shock protein 90kDa alpha (cytosolic), class B member 1; heat shock protein beta; Heat shock protein HSP 90-beta; HS90B; HSP 84; HSP 90; HSP84; HSP90-BETA; HSP90AB1; HSP90B; HSPC2; HSPCB

Product Type Specific: HSP90AB1 phosphosite-specific antibody

Antibody Code: PN520

Antibody Target Type: Phosphosite-specific

Antibody Phosphosite: Y484

Protein UniProt: P08238

Protein SigNET: HSP90AB1

Antibody Type: Polyclonal

Antibody Host Species: Rabbit

Antibody Immunogen Source: Human HSP90AB1 (HSP90B; HSP90-beta) sequence peptide Cat. No.: PE-04AFE99

Antibody Immunogen Sequence: KSI(pY)YIT(bA)C

Antibody Immunogen Description: Corresponds to amino acid residues K481 to T487; in the middle of the HSP90 domain. The effect of Y484 phosphorylation is unclear. This is the major in vivo phosphorylation site in HSP90AB1 (≥814 reports from high throughput mass spectrometry studies recorded in PhosphoSitePlus).

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 with immunogen peptide on dot blots.

Antibody Positive Control: The observed molecular mass of the processed target protein on SDS-PAGE gels is reported to be around 90-95 kDa.

Antibody Specificity: High-very high

Antibody Cross Reactivity: No significant cross-reactive proteins detected in A431 and Jurkat cells.

Scientific Background: HSP90 is an abundantly and ubiquitously expressed heat shock protein. It is understood to exist in two principal forms α and β, which share 85% sequence amino acid homology. The two isoforms of HSP90 are expressed in the cytosolic compartment (1). Despite the similarities, HSP90α exists predominantly as a homodimer while HSP90β exists mainly as a monomer (2). They feature three main domains: an amino-terminal domain (NTD) for ATP binding, a middle domain (MD) for client protein binding, and a carboxy-terminal domain (CTD) for dimerization. From a functional perspective, HSP90 participates in the folding, assembly, maturation, and stabilization of specific proteins as an integral component of a chaperone complex (3-6). Furthermore, HSP90 is highly conserved between species; having 60% and 78% amino acid similarity between mammalian and the corresponding yeast and Drosophila proteins, respectively. Despite its label of being a heat-shock protein, HSP90 is one of the most highly expressed proteins in unstressed cells (1–2% of cytosolic protein). It carries out a number of housekeeping functions – including controlling the activity, turnover, and trafficking of a variety of proteins. Most of the HSP90-regulated proteins that have been discovered to date are involved in cell signalling (7-8). The number of identified proteins known to interact with HSP90 is over 100. Target proteins include the kinases v-Src, Wee1, and c-Raf, transcriptional regulators such as p53 and steroid receptors, and the polymerases of the hepatitis B virus and telomerase (5). When bound to ATP, HSP90 interacts with co-chaperones Cdc37, p23, and an assortment of immunophilin-like proteins, forming a complex that stabilizes and protects target proteins from proteasomal degradation. In most cases, HSP90-interacting proteins have been shown to co-precipitate with HSP90 when carrying out immunoadsorption studies, and to exist in cytosolic heterocomplexes with it. In a number of cases, variations in HSP90 expression or HSP90 mutation has been shown to degrade signalling function via the protein or to impair a specific function of the protein (such as steroid binding, kinase activity) in vivo. Its activity is regulated by numerous co-chaperones (such as p60/Hop, p50Cdc37, p23) that assist in the chaperoning process. Ansamycin antibiotics, such as geldanamycin and radicicol, inhibit HSP90 function (9). HSP90 is essential for cancer cell survival by stabilizing mutated or overexpressed proteins. This description may include information annotated by UniProt and/or Google AI.

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