Overview: Product Name HSP90 Antibody DescriptionMouse Anti-Human HSP90 Monoclonal IgG2a Species Reactivity Dog, Human, Mouse, Rat, Chicken, Fish, Gummy Shark (Mustelus antarcticus), Rabbit, School Shark (Galeorhinus galeus), Shark, White Sucker Fish (Catostomus commersonii) Applications WB, IHC, ICC/IF, IP, ELISA, AM Antibody Dilution WB (1:2500), IHC (1:100); optimal dilutions for assays should be determined by the user. Host Species Mouse Immunogen Species Human Immunogen Recombinant human HSP90beta 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 H9010 Isotype IgG2a Specificity Detects 90kDa. Detects HSP90 beta in all reactive species except in Chicken, where it detects both alpha and beta isoforms. Cite This Product StressMarq Biosciences Cat# SMC-107, RRID: AB_854214 Certificate of Analysis 1 µg/ml of SMC-107 was sufficient for detection of HSP90beta 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 HSP84 Antibody, HSP90 Antibody, HSP90 beta Antibody, HSP90B Antibody, HSPC2 Antibody, HSPCB Antibody Research Areas Cancer, Heat Shock Cellular Localization Cytoplasm, Melanosome Accession Number NP_031381.2 Gene ID 3326 Swiss Prot P08238 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 S.J.H., 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 W.B. (1998) Proc Soc Exptl Biol Med 217: 420–434. 9. Whitesell L., et al. (1994) Proc Natl Acad Sci USA 91: 8324–8328. 10. Barent R. L. (1998) Mol. Endocrinol. 12: 342-354 11. Lo. M.A. (1998) EMBO J. 17: 6879-6887. Product Images Immunohistochemistry analysis using Mouse Anti-Hsp90 Monoclonal Antibody, Clone H9010 (SMC-107). Tissue: inflamed colon. Species: Mouse. Fixation: Formalin. Primary Antibody: Mouse Anti-Hsp90 Monoclonal Antibody (SMC-107) at 1:10000 for 12 hours at 4°C. Secondary Antibody: Alexa Fluor 555 Goat Anti-Mouse (red) at 1:5000 for 1 hour at RT. Localization: Inflammatory and epithelial mucosa. Magnification: 40x. Immunohistochemistry analysis using Mouse Anti-Hsp90 Monoclonal Antibody, Clone H9010 (SMC-107). Tissue: colon carcinoma. Species: Human. Fixation: Formalin. Primary Antibody: Mouse Anti-Hsp90 Monoclonal Antibody (SMC-107) at 1:10000 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 Human cell lysates from various cell lines showing detection of Hsp90 protein using Mouse Anti-Hsp90 Monoclonal Antibody, Clone H9010 (SMC-107). Load: 15 µg protein. Block: 1.5% BSA for 30 minutes at RT. Primary Antibody: Mouse Anti-Hsp90 Monoclonal Antibody (SMC-107) 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 Monoclonal Antibody, Clone H9010 (SMC-107). Tissue: colon carcinoma. Species: Human. Fixation: Formalin. Primary Antibody: Mouse Anti-Hsp90 Monoclonal Antibody (SMC-107) at 1:10000 for 12 hours at 4°C. Secondary Antibody: Alexa Fluor 555 Goat Anti-Mouse (red) at 1:5000 for 1 hour at RT. Magnification: 40x. Immunohistochemistry analysis using Mouse Anti-Hsp90 Monoclonal Antibody, Clone H9010 (SMC-107). Tissue: inflamed colon. Species: Mouse. Fixation: Formalin. Primary Antibody: Mouse Anti-Hsp90 Monoclonal Antibody (SMC-107) at 1:10000 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. Immunohistochemistry analysis using Mouse Anti-Hsp90 Monoclonal Antibody, Clone H9010 (SMC-107). Tissue: backskin. Species: Mouse. Fixation: Bouin’s Fixative and paraffin-embedded. Primary Antibody: Mouse Anti-Hsp90 Monoclonal Antibody (SMC-107) at 1:100 for 1 hour at RT. Secondary Antibody: FITC Goat Anti-Mouse (green) at 1:50 for 1 hour at RT. Localization: Epidermis. Western Blot analysis of Human HeLa cell lysates showing detection of Hsp90 protein using Mouse Anti-Hsp90 Monoclonal Antibody, Clone H9010 (SMC-107). Primary Antibody: Mouse Anti-Hsp90 Monoclonal Antibody (SMC-107) at 1:1000. Secondary Antibody: HRP Goat Anti-Mouse. Western blot analysis of Human Lysates showing detection of Hsp90 protein using Mouse Anti-Hsp90 Monoclonal Antibody, Clone H9010 (SMC-107). Primary Antibody: Mouse Anti-Hsp90 Monoclonal Antibody (SMC-107) at 1:1000. Comparison of clone H9010 behavior with Hsp90 human beta (1) and Hsp90 human alpha (2). Courtesy of: David Toft, Mayo Clinic. Western blot analysis of Human Lysates showing detection of Hsp90 protein using Mouse Anti-Hsp90 Monoclonal Antibody, Clone H9010 (SMC-107). Primary Antibody: Mouse Anti-Hsp90 Monoclonal Antibody (SMC-107) at 1:1000. Comparison of clone H9010 behavior with Hsp90 human beta (1) and Hsp90 human alpha (2). Courtesy of: David Toft, Mayo Clinic. Product Citations (24) Western Blot PRAS40 Connects Microenvironmental Stress Signaling to Exosome-Mediated Secretion. Guo, J. et al. (2017) Mol Cell Biol. [Epub ahead of print]. PubMed ID: 28674187 Reactivity Human Applications: Western Blot Extracellular heat shock protein 90α mediates HDM-induced bronchial epithelial barrier dysfunction by activating RhoA/MLC signaling. Dong, H.M. et al. (2017) Respir Res. 18(1):111. PubMed ID: 28558721 Reactivity Human Applications: Western Blot Extracellular Hsp90 and TGFβ regulate adhesion, migration and anchorage independent growth in a paired colon cancer cell line model. de la Mare, J.A., Jurgens, T. and Edkins, A.L. (2017) BMC Cancer. 17(1):202. PubMed ID: 28302086 Reactivity Human Applications: Western Blot Other Citations Characterization and Functional Analysis of Extracellular Vesicles and Muscle-Abundant miRNAs (miR-1, miR-133a, and miR-206) in C2C12 Myocytes and mdx Mice. Matsuzaka, Y. et al. (2016) PLoS One. 11(12):e0167811. PubMed ID: 27977725 Reactivity Mouse Applications: Western blot Evolutionarily conserved dual lysine motif determines the non-chaperone function of secreted Hsp90alpha in tumour progression. Zou, M. et al. (2016) Oncogene. 36(15):2160-2171. PubMed ID: 27721406 Reactivity Human Applications: Western Blot The epichaperome is an integrated chaperome network that facilitates tumour survival. Rodina, A. et al. (2016) Nature. 538(7625):397-401. PubMed ID: 27706135 Reactivity Human Applications: Western Blot Physiological responses to hypersalinity correspond to nursery ground usage in two inshore shark species (Mustelus antarcticus & Galeorhinus galeus). Tunnah, L. et al. (2016) J Exp Biol. 219(Pt 13):2028-38. PubMed ID: 27207636 Reactivity Shark Applications: Western Blot COMPOSITIONS AND METHODS FOR TREATING HIF-1a OVER-EXPRESSING CANCERS. Li, W. et al. (2016) United States Patent Application 20160053003 PubMed ID: N/A Reactivity Human Applications: Western Blot Breast Cancer MDA-MB-231 Cells Use Secreted Heat Shock Protein-90alpha (Hsp90α) to Survive a Hostile Hypoxic Environment. Dong, H. et al. (2016) Sci Rep. 6:20605. PubMed ID: 26846992 Reactivity Human Applications: Western Blot Pharmacoproteomics identifies combinatorial therapy targets for diffuse large B cell lymphoma. Goldstein, R.L. et al. (2015) J Clin Invest. 2015. pii: 80714 PubMed ID: 26529251 Reactivity Human Applications: Western Blot Cold acclimation increases levels of some heat shock protein and sirtuin isoforms in threespine stickleback. Teigen, L.E., Orczewska, J.I., McLaughlin, J. (2015) Comp Biochem Physiol A Mol Integr Physiol. 188:139-47. PubMed ID: 26123780 Reactivity Gasterosteus aculeatus (Three-spined stickleback) Applications: Western Blot Hsp90α and Hsp90β together operate a hypoxia and nutrient paucity stress-response mechanism during wound healing. Jayaprakash, P. et al. (2015) J Cell Sci. 128(8):1475-80. PubMed ID: 25736295 Reactivity Human Applications: Western Blot Cardiorespiratory toxicity of environmentally relevant zinc oxide nanoparticles in the freshwater fish Catostomus commersonii. Bessemer, R.A. et al. (2014) Nanotoxicology. 27:1-10. PubMed ID: 25427894 Reactivity Catostomus commersonii (fish) Applications: Western Blot Hsp90 Binds Directly to Fibronectin (FN) and Inhibition Reduces the Extracellular Fibronectin Matrix in Breast Cancer Cells. Hunter, M. C. et al. (2014) PLoS One. 9(1):e86842. PubMed ID: 24466266 Reactivity Human Applications: Western Blot Uses of Labeled HSP90 Inhibitors. Chiosis, G. et al. (2014) United States Patent Application 20140294725 Kind Code: A1 PubMed ID: N/A Reactivity Human Applications: Western Blot Paralog-selective Hsp90 inhibitors define tumor-specific regulation of HER 2. Patel, P.D. et al. (2013) Nat Chem Biol. 9(11):677-84. PubMed ID: 23995768 Reactivity Human Applications: Western Blot A Novel Neurotrophic Drug for Cognitive Enhancement and Alzheimer's Disease. Chen, Q. et al. (2011) PLoS One. 6 (12): e27865. PubMed ID: 22194796 Reactivity Rat Applications: Western Blot Design, synthesis, and evaluation of small molecule Hsp90 probes. Taldone, T. et al. (2011) Bioorg Med Chem. 19 (8): 2603-2614. PubMed ID: 21459002 Reactivity Human Applications: Western Blot Key motifs in EBV (Epstein-Barr virus)-encoded protein kinase for phosphorylation activity and nuclear localization. Gershburg, S., Murphy, L., Marschall, M. and Gershburg, E. (2010) Biochem J. 431 (2): 227-235. PubMed ID: 20704565 Reactivity Human Applications: Western Blot The epichaperome is an integrated chaperome network that facilitates tumour survival. Rodina, A. et al. (2016) Nature. 538(7625):397-401. PubMed ID: 27706135 Reactivity Human Applications: Immunocytochemistry/Immunofluorescence Breast Cancer MDA-MB-231 Cells Use Secreted Heat Shock Protein-90alpha (Hsp90α) to Survive a Hostile Hypoxic Environment. Dong, H. et al. (2016) Sci Rep. 6:20605. PubMed ID: 26846992 Reactivity Human Applications: Immunocytochemistry/Immunofluorescence Biomarker Analysis with Grating Coupled Surface Plasmon Coupled Fluorescence. Mendoza, A., Dias, J.A., Zeltner, T. and Lawrence, D.A. (2014) J Adv Bio & Biotech. 1(1): 1-22. PubMed ID: N/A Reactivity Human Applications: Antibody Microarray Biomarker Analysis with Grating Coupled Surface Plasmon Coupled Fluorescence. Mendoza, A., Dias, J.A., Zeltner, T. and Lawrence, D.A. (2014) J Adv Bio & Biotech. 1(1): 1-22. PubMed ID: N/A Reactivity Mouse Applications: Antibody Microarray HDAC6 Regulates Glucocorticoid Receptor Signaling in Serotonin Pathways with Critical Impact on Stress Resilience. Espallergues, J. et al. (2012) J Neurosci. 32 (13): 4400-4416. PubMed ID: 22457490 Reactivity Mouse Applications: ImmunoprecipitationStreptavidinProperties: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 Datasheet