Heat Shock Protein 90 alpha (HSP90 alpha) (-alpha acetyl specific K294) Antikörper

Antigen: Heat Shock Protein 90 alpha (HSP90 alpha)

Synonyme: HSP90AA1, HSPCA

Epitope: -alpha acetyl specific K294

Klonalität: Polyklonal

Wirt: Kaninchen

Reaktivität: Human, Rind (Kuh), Maus, Ratte (Rattus)

Applikation: ELISA, Western Blot (WB), Immunpräzipitation (IP)

Produktnummer: ABIN233817

Produktbeschreibung

Weitere Bezeichnung: Heat Shock Protein HSP 90-alpha acetyl specific K294

Swiss-Prot: P46633

Immunogen: This affinity purified antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to amino acids surrounding K294 of human Hsp90.

Format: Liquid (sterile filtered)

Beschreibung: Hsp90 is a member of the heat shock protein 90 family, the members of which are highly conserved between isoforms and species. Hsp90 functions as a molecular chaperone and has ATPase activity. Hsp90 is a cytoplasmic protein that forms a homodimer in vivo and interacts with TOM34, AHSA1, HDAC6 and SMYD3. Several signal transduction pathways depend on Hsp90 function, including pathways involving erbB2, hypoxia sensitivity (Hif1 alpha), and steroid hormone receptors (for example, androgen, progesterone, glucocorticoid, and aryl-hydrocarbon). Recent reports show that Hsp90 from tumor cells has increased sensitivity to small molecule inhibitors (for example, 17AAG). The mechanism of the differential sensitivity of Hsp90 from normal versus tumor cells is unknown, although mutation has been ruled out. One possible mechanism may be differences in post-translational modification of tumor Hsp90. K294 was found to be acetylated in purified Hsp90 from SkBr3 cells, a breast cancer cell line.

Spezifität: This affinity purified antibody is directed against human Hsp90 protein acetylated at K294. The product was affinity purified from monospecific antiserum by immunoaffinity chromatography using acetyl-peptide coupled to agarose beads followed by solid phase adsorption against non-acetyl peptide. While ELISA data show strong reactivity with the acetylated form of the immunizing peptide and minimal reactivity with the non-acetylated form, to date western blotting data are not definitive for acetyl K294 specificity as blots show equivalent staining of lysates from cells either treated or untreated with Trichostatin A (an HDAC inhibitor). A BLAST analysis was used to suggest cross-reactivity with Hsp90 from human, mouse, rat, monkey, chicken and Drosophila based on 100% homology with the immunizing sequence. Reactivity of this antibody with Hsp90 from other species is unknown.

Anwendungen

Applikationshinweise: Recommended Dilutions: ELISA 1:2,500 - 1:10,000 WESTERN BLOT 1:500 - 1:2,500 IMMUNOHISTOCHEMISTRY 5-10 ug/ml OTHER APPLICATIONS User Optimized. Note: This affinity purified antibody has been tested for use in ELISA, immunohistochemistry and western blot. This antibody reacts strongly with the acetylated form of the immunizing peptide and shows minimal reactivity with the non-acetylated form when tested by ELISA. To date, western blotting shows equivalent staining of lysates either treated or untreated with Trichostatin A (an HDAC inhibitor). Therefore, western blotting results are not definitive for demonstrating the specificity of this reagent. Specific conditions for reactivity should be optimized by the end user. Expect a band approximately 90 kDa in size corresponding to Hsp90 protein by western blotting in the appropriate cell lysate or extract.

Konzentration: 0.96 mg/ml (by UV absorbance at 280 nm)

Buffer: Stabilizer: None. Preservative: 0.01% (w/v) Sodium Azide. Buffer: 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2.

Lagerung: Store vial at -20° C prior to opening. Dilute only prior to immediate use. For extended storage, aliquot contents and freeze at -20° C or below. Avoid cycles of freezing and thawing.

Forschungsgebiet: Hitzeschockproteine, Krebs

Beschränkungen: Nur für Forschungszwecke einsetzbar

Bilder

Immunohistochemistry. This affinity purified anti-Hsp90 acetyl K294 antibody was used at 20 μg/ml to detect signal in a variety of tissues including multi-human, multi-brain and multi-cancer slides. This image shows moderate nuclear and granular cytoplasmic positive staining in human prostate carcinoma at 40X. Tissue was formalin-fixed and paraffin embedded. The image shows localization of the antibody as the precipitated red signal, with a hematoxylin purple nuclear counterstain.

Publikationen

Publikationen: Dedes, Dedes, Imesch et al.: "Acquired vorinostat resistance shows partial cross-resistance to 'second-generation' HDAC inhibitors and correlates with loss of histone acetylation and apoptosis but not with altered HDAC and HAT activities." in: Anti-cancer drugs, Vol. 20, Issue 5, pp. 321-33, 2009 (PubMed).