Overview: Product Name HSP90 alpha Antibody DescriptionMouse Anti-Human HSP90 alpha Monoclonal IgG2a Species Reactivity Human, Mouse, Rat Applications WB, IHC, ELISA Antibody Dilution WB (1:1000), IHC (1:5000); optimal dilutions for assays should be determined by the user. Host Species Mouse Immunogen Species Human Immunogen Recombinant human HSP90alpha; Specificity mapped to amino acids 604-731 Concentration 1 mg/ml Conjugates Alkaline Phosphatase, APC, ATTO 390, ATTO 488, ATTO 565, ATTO 594, ATTO 633, ATTO 655, ATTO 680, ATTO 700, Biotin, FITC, HRP, PE/ATTO 594, PerCP, RPE, Streptavidin, Unconjugated PerCP Overview:Peridinin-Chlorophyll-Protein ComplexSmall phycobiliproteinIsolated from red algaeLarge stokes shift (195 nm)Molecular Weight: 35 kDaPerCP Datasheet Optical Properties:λex = 482 nmλem = 677 nmεmax = 1.96 x 106Laser = 488 nm  PE/ATTO 594PE/ATTO 594 is a tandem conjugate, where PE is excited at 535 nm and transfers energy to ATTO 594 via FRET (fluorescence resonance energy transfer), which emits at 627 nm.Overview:High fluorescence yieldHigh photostabilityVery hydrophilicExcellent solubility in waterVery little aggregationPE/ATTO 594 DatasheetOptical Properties:λex = 535 nmλem = 627 nmLaser = 488 to 561 nm   FITC (Fluorescein)Overview:Excellent fluorescence quantum yieldHigh rate of photobleachingGood solubility in waterBroad emission spectrumpH dependent spectraMolecular formula: C20H12O5Molar mass: 332.3 g/molFITC-Fluorescent-conjugateOptical Properties:λex = 494 nmλem = 520 nmεmax = 7.3×104Φf = 0.92τfl = 5.0 nsBrightness = 67.2Laser = 488 nmFilter set = FITC  ATTO 700Overview:High fluorescence yieldExcellent thermal and photostabilityQuenched by electron donorsVery hydrophilicGood solubility in polar solventsZwitterionic dyeMolar Mass: 575 g/molATTO 700 Datasheet Optical Properties:λex = 700 nmλem = 719 nmεmax = 1.25×105Φf = 0.25τfl = 1.6 nsBrightness = 31.3Laser = 676 nmFilter set = Cy®5.5  ATTO 680Overview:High fluorescence yieldExcellent thermal and photostabilityQuenched by electron donorsVery hydrophilicGood solubility in polar solventsZwitterionic dyeMolar Mass: 631 g/molATTO 680 Datasheet Optical Properties:λex = 680 nmλem = 700 nmεmax = 1.25×105Φf = 0.30τfl = 1.7 nsBrightness = 37.5Laser = 633 – 676 nmFilter set = Cy®5.5  ATTO 655Overview:High fluorescence yieldHigh thermal and photostabilityExcellent ozone resistanceQuenched by electron donorsVery hydrophilicGood solubility in polar solventsZwitterionic dyeMolar Mass: 634 g/molATTO 655 DatasheetOptical Properties:λex = 663 nmλem = 684 nmεmax = 1.25×105Φf = 0.30τfl = 1.8 nsBrightness = 37.5Laser = 633 – 647 nmFilter set = Cy®5  ATTO 633Overview:High fluorescence yieldHigh thermal and photostabilityModerately hydrophilicGood solubility in polar solventsStable at pH 4 – 11Cationic dye, perchlorate saltMolar Mass: 652.2 g/molATTO 633 DatasheetOptical Properties:λex = 629 nmλem = 657 nmεmax = 1.3×105Φf = 0.64τfl = 3.2 nsBrightness = 83.2Laser = 633 nmFilter set = Cy®5  ATTO 594Overview:High fluorescence yieldHigh photostabilityVery hydrophilicExcellent solubility in waterVery little aggregationNew dye with net charge of -1Molar Mass: 1137 g/molATTO 594 Datasheet Optical Properties:λex = 601 nmλem = 627 nmεmax = 1.2×105Φf = 0.85τfl = 3.5 nsBrightness = 102Laser = 594 nmFilter set = Texas Red®  ATTO 565Overview:High fluorescence yieldHigh thermal and photostabilityGood solubility in polar solventsExcellent solubility in waterVery little aggregationRhodamine dye derivativeMolar Mass: 611 g/molATTO 565 Datasheet Optical Properties:λex = 563 nmλem = 592 nmεmax = 1.2×105Φf = 0.9τfl = 3.4 nBrightness = 10Laser = 532 nmFilter set = TRITC   ATTO 488Overview:High fluorescence yieldHigh photostabilityVery hydrophilicExcellent solubility in waterVery little aggregationNew dye with net charge of -1Molar Mass: 804 g/mol ATTO 488 Datasheet  Optical Properties:λex = 501 nmλem = 523 nmεmax = 9.0×104Φf = 0.80τfl = 4.1 nsBrightness = 72Laser = 488 nmFilter set = FITC   ATTO 390Overview:High fluorescence yieldLarge Stokes-shift (89 nm)Good photostabilityModerately hydrophilicGood solubility in polar solventsCoumarin derivate, unchargedLow molar mass: 343.42 g/mol ATTO 390 DatasheetOptical Properties:λex = 390 nmλem = 479 nmεmax = 2.4×104Φf = 0.90τfl = 5.0 nsBrightness = 21.6Laser = 365 or 405 nm APC (Allophycocyanin)Overview:High quantum yieldLarge phycobiliprotein6 chromophores per moleculeIsolated from red algaeMolecular Weight: 105 kDaAPC Datasheet Optical Properties:λex = 650 nmλem = 660 nmεmax = 7.0×105Φf = 0.68Brightness = 476Laser = 594 or 633 nmFilter set = Cy®5 StreptavidinProperties:Homo-tetrameric protein purified from Streptomyces avidinii which binds four biotin molecules with extremely high affinityMolecular weight: 53 kDaFormula: C10H16N2O3SApplications: Western blot, immunohistochemistry, and ELISAStreptavidin DatasheetBiotinProperties:Binds tetrameric avidin proteins including Streptavidin and neuravidin with very high affinityMolar mass: 244.31 g/molFormula: C10H16N2O3SApplications: Western blot, immunohistochemistry, and ELISABiotin DatasheetHRP (Horseradish peroxidase)Properties:Enzymatic activity is used to amplify weak signals and increase visibility of a targetReadily combines with hydrogen peroxide (H2O2) to form HRP-H2O2 complex which can oxidize various hydrogen donorsCatalyzes the conversion of:Chromogenic substrates (e.g. TMB, DAB, ABTS) into coloured productsChemiluminescent substrates (e.g. luminol and isoluminol) into light emitting products via enhanced chemiluminescence (ECL)Fluorogenic substrates (e.g. tyramine, homovanillic acid, and 4-hydroxyphenyl acetic acid) into fluorescent productsHigh turnover rate enables rapid generation of a strong signal44 kDa glycoproteinExtinction coefficient: 100 (403 nm)Applications: Western blot, immunohistochemistry, and ELISAHRP DatasheetAP (Alkaline Phosphatase)Properties:Broad enzymatic activity for phosphate esters of alcohols, amines, pyrophosphate, and phenolsCommonly used to dephosphorylate the 5’-termini of DNA and RNA to prevent self-ligationCatalyzes the conversion of:Chromogenic substrates (e.g. pNPP, naphthol AS-TR phosphate, BCIP) into coloured productsFluorogenic substrates (e.g. 4-methylumbelliferyl phosphate) into fluorescent productsMolecular weight: 140 kDaApplications: Western blot, immunohistochemistry, and ELISAAP Datasheet  R-PE (R-Phycoerythrin)Overview:Broad excitation spectrumHigh quantum yieldPhotostableMember of the phycobiliprotein familyIsolated from red algaeExcellent solubility in waterMolecular Weight: 250 kDaR-PE Datasheet Optical Properties:λex = 565 nmλem = 575 nmεmax = 2.0×106 Φf = 0.84Brightness = 1.68 x 103Laser = 488 to 561 nmFilter set = TRITC  Properties Storage Buffer PBS pH7.2, 50% glycerol, 0.09% sodium azide Storage Temperature -20ºC Shipping Temperature Blue Ice or 4ºC Purification Protein G Purified Clonality Monoclonal Clone Number Hyb-K41009 Isotype IgG2a Specificity Detects 90kDa. This is an alpha-specific product. Cite This Product StressMarq Biosciences Cat# SMC-108, RRID: AB_854218 Certificate of Analysis 1 µg/ml of SMC-108 was sufficient for detection of HSP90alpha in 20 µg of heat shocked HeLa cell lysate by colorimetric immunoblot analysis using Goat anti-mouse IgG:HRP as the secondary antibody. Biological Description Alternative Names HSP86 Antibody, HSP89A Antibody, HSP90A Antibody, HSP90AA1 Antibody, HSP90Alpha Antibody, HSPC1 Antibody, HSPCA Antibody, HSPCAL3 Antibody Research Areas Cancer, Heat Shock Cellular Localization Cytoplasm, Melanosome Accession Number NP_001017963.2 Gene ID 3320 Swiss Prot P07900 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). 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. HSP90 is a highly conserved and essential stress protein that is expressed in all eukaryotic cells. Despite it’s 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 signaling (7-8). The number of proteins now know to interact with HSP90 is about 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 signaling function via the protein or to impair a specific function of the protein (such as steroid binding, kinase activity) in vivo. Ansamycin antibiotics, such as geldanamycin and radicicol, inhibit HSP90 function (9). For more information visit our HSP90 Scientific Resource Guide at http://www.HSP90.ca. References 1. Nemoto, T. et al. (1997) J.Biol Chem. 272: 26179-26187. 2. Minami, Y, et al. (1991), J.Biol Chem. 266: 10099-10103. 3. Arlander SJH, et al. (2003) J Biol Chem. 278: 52572-52577. 4. Pearl H, et al. (2001) Adv Protein Chem. 59: 157-186. 5. Neckers L, et al. (2002) Trends Mol Med. 8: S55-S61. 6. Pratt W, Toft D. (2003) Exp Biol Med. 228: 111-133. 7. Pratt W, Toft D. (1997) Endocr Rev. 18: 306–360. 8. Pratt WB. (1998) Proc Soc Exptl Biol Med. 217: 420–434. 9. Whitesell L, et al. (1994) Proc Natl Acad Sci USA. 91: 8324–8328. 10. Nemoto, T. (1997) Biochem and Mol. Bio Intl. 42 (5): 881-889. Product Images Immunohistochemistry analysis using Mouse Anti-Hsp90 alpha Monoclonal Antibody, Clone K41009 (SMC-108). Tissue: colon carcinoma. Species: Human. Fixation: Formalin. Primary Antibody: Mouse Anti-Hsp90 alpha Monoclonal Antibody (SMC-108) at 1:5000 for 12 hours at 4°C. Secondary Antibody: Biotin Goat Anti-Mouse at 1:2000 for 1 hour at RT. Counterstain: Mayer Hematoxylin (purple/blue) nuclear stain at 200 µl for 2 minutes at RT. Localization: Inflammatory cells. Magnification: 40x. Western Blot analysis of Rat Lysates showing detection of Hsp90 alpha protein using Mouse Anti-Hsp90 alpha Monoclonal Antibody, Clone K41009 (SMC-108). Load: 15 µg protein. Block: 1.5% BSA for 30 minutes at RT. Primary Antibody: Mouse Anti-Hsp90 alpha Monoclonal Antibody (SMC-108) at 1:1000 for 2 hours at RT. Secondary Antibody: Sheep Anti-Mouse IgG: HRP for 1 hour at RT. Immunohistochemistry analysis using Mouse Anti-Hsp90 alpha Monoclonal Antibody, Clone K41009 (SMC-108). Tissue: inflamed colon. Species: Mouse. Fixation: Formalin. Primary Antibody: Mouse Anti-Hsp90 alpha Monoclonal Antibody (SMC-108) at 1:5000 for 12 hours at 4°C. Secondary Antibody: Biotin Goat Anti-Mouse at 1:2000 for 1 hour at RT. Counterstain: Mayer Hematoxylin (purple/blue) nuclear stain at 200 µl for 2 minutes at RT. Localization: Inflammatory cells. Magnification: 40x. Product Citations (7) Western Blot Heat shock protein 90 ensures efficient mumps virus replication by assisting with viral polymerase complex formation Katoh, H. et al. (2017) J Virol. pii: JVI.02220-16. PubMed ID: 28053100 Reactivity Human Applications: Western Blot Effects of Long-Term Exposure to 60 GHz Millimeter-Wavelength Radiation on the Genotoxicity and Heat Shock Protein (Hsp) Expression of Cells Derived from Human Eye. Koyama, S. et al. (2016) Int J Environ Res Public Health. 13(8). pii: E802. PubMed ID: 27509516 Reactivity Human Applications: Western Blot Twenty Four-Hour Exposure to a 0.12 THz Electromagnetic Field Does Not Affect the Genotoxicity, Morphological Changes, or Expression of Heat Shock Protein in HCE-T Cells. Koyama, S. et al. (2016) Int J Environ Res Public Health. 13(8). pii: E802. PubMed ID: 27527204 Reactivity Human Applications: Western Blot Heat-induced expression of the immediate-early gene IER5 and its involvement in the proliferation of heat-shocked cells. Ishikawa, Y. and Sakurai, H. (2014) FEBS J. 282(2):332-40. PubMed ID: 25355627 Reactivity Human Applications: Western Blot Heat Shock Factor Hsf1 Cooperates with ErbB2 (Her2/Neu) Protein to Promote Mammary Tumorigenesis and Metastasis. Xi, C., Hu, Y., Buckhaults, P., Moskophidis, D. and Mivechi, N.F. (2012) J Biol Chem. 287, 35646-35657. PubMed ID: 22847003 Reactivity Human Applications: Western Blot Heat Shock Protein 90α (Hsp90α) Is Phosphorylated in Response to DNA Damage and Accumulates in Repair Foci. Quanz, M. et al. (2012) J Biol Chem. 287, 8803-8815. PubMed ID: 22270370 Reactivity Human Applications: Western Blot Other Citations Associations of HSP90 Client Proteins in Human Breast Cancer. Shipp, C., Watson, K., and Jones, G.L. (2011) Anticancer Research. 31 (6): 2095-2101. PubMed ID: 21737627 Reactivity Human Applications: Immunoprecipitation  ATTO 488Overview:High fluorescence yieldHigh photostabilityVery hydrophilicExcellent solubility in waterVery little aggregationNew dye with net charge of -1Molar Mass: 804 g/mol ATTO 488 Datasheet  Optical Properties:λex = 501 nmλem = 523 nmεmax = 9.0×104Φf = 0.80τfl = 4.1 nsBrightness = 72Laser = 488 nmFilter set = FITC