Overview: Product Name HIF2 alpha Antibody DescriptionMouse Anti-Human HIF2 alpha Monoclonal IgG1 Species Reactivity Human, Mouse Applications WB, IHC, ICC/IF, ELISA, AM Antibody Dilution WB (1:500), IHC (1:100), ICC/IF (1:100); optimal dilutions for assays should be determined by the user. Host Species Mouse Immunogen Species Human Immunogen GST-human EPAS-1 (HIF2alpha) amino acids 535-631 fusion protein 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 pH7.4, 50% glycerol, 0.09% sodium azide Storage Temperature -20ºC Shipping Temperature Blue Ice or 4ºC Purification Protein G Purified Clonality Monoclonal Clone Number EP190B Isotype IgG1 Specificity Detects ~100kDa. Specific for HIF2 Alpha. Cite This Product StressMarq Biosciences Cat# SMC-185, RRID: AB_10643260 Certificate of Analysis 1 µg/ml of SMC-185 was sufficient for detection of HIF2α in 20 µg of CoCl2-induced Hela cell lysate by colorimetric immunoblot analysis using Goat anti-mouse IgG:HRP as the secondary antibody. Biological Description Alternative Names ECYT4 Antibody, EPAS1 Antibody, HIF2alpha Antibody, HIF1 Alpha like factor Antibody, HLF Antibody, Hypoxia inducible factor 2 alpha Antibody, MOP2 Antibody, PASD2 Antibody Research Areas Cancer, Cell Signaling, Epigenetics, Epigenetics and Nuclear Signaling, Oxidative Stress Cellular Localization Nucleus Accession Number NP_001421.4 Gene ID 2034 Swiss Prot Q99814 Scientific Background Members of the hypoxia-inducible factor (HIF) family of transcription factors regulate the cellular response to hypoxia (1). HIF2-Alpha is involved in catecholamine homeostasis, vascular remodeling, physiological angiogenesis and adipogenesis. It is overexpressed in many cancerous tissues, but its exact role in tumourprogression remains to be clarified (2). Studies suggest that in the case of non-small cell lung cancer, HIF2-Alpha actually is a promoter of tumor growth and progression in a solid tumor (1). Other data suggests that HIF2-Alpha is an important regulator of innate immunity, and therefore may be useful in the therapeutic target for treating inflammatory disorders and cancer (3). References 1. Kim W.Y., et al. (2009) J Clin Invest. 119(8): 2160-2170. 2. Favier J., Lapointe S., Maliba R. and Sirois M.G. (2007) BMC Cancer. 7:139. 3. Imtiyaz H.Z., et al. (2010) J Clin Invest. 120(8): 2699-2714. Product Images Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-HIF2 alpha Monoclonal Antibody, Clone EP190b (SMC-185). Tissue: HeLa Cells. Species: Human. Fixation: 2% Formaldehyde for 20 min at RT. Primary Antibody: Mouse Anti-HIF2 alpha Monoclonal Antibody (SMC-185) at 1:100 for 12 hours at 4°C. Secondary Antibody: APC Goat Anti-Mouse (red) at 1:200 for 2 hours at RT. Counterstain: DAPI (blue) nuclear stain at 1:40000 for 2 hours at RT. Localization: Cytoplasm. Localizes to the nucleus upon hypoxia. Magnification: 100x. (A) DAPI (blue) nuclear stain. (B) Anti-HIF2 alpha Antibody. (C) Composite. Western Blot analysis of Human HeLa cell lysates showing detection of HIF2 alpha protein using Mouse Anti-HIF2 alpha Monoclonal Antibody, Clone EP190b (SMC-185). Load: 15 µg protein. Block: 1.5% BSA for 30 minutes at RT. Primary Antibody: Mouse Anti-HIF2 alpha Monoclonal Antibody (SMC-185) at 1:1000 for 2 hours at RT. Secondary Antibody: Sheep Anti-Mouse IgG: HRP for 1 hour at RT. Cells CoCl treated. Immunohistochemistry analysis using Mouse Anti-HIF2 alpha Monoclonal Antibody, Clone EP190b (SMC-185). Tissue: backskin. Species: Mouse. Fixation: Bouin’s Fixative and paraffin-embedded. Primary Antibody: Mouse Anti-HIF2 alpha Monoclonal Antibody (SMC-185) at 1:100 for 1 hour at RT. Secondary Antibody: FITC Goat Anti-Mouse (green) at 1:50 for 1 hour at RT. Localization: Basal cell staining in the epidermis, dermis positive and muscle positive. . Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-HIF2 alpha Monoclonal Antibody, Clone EP190b (SMC-185). Tissue: HeLa Cells. Species: Human. Fixation: 2% Formaldehyde for 20 min at RT. Primary Antibody: Mouse Anti-HIF2 alpha Monoclonal Antibody (SMC-185) at 1:100 for 12 hours at 4°C. Secondary Antibody: FITC Goat Anti-Mouse (green) at 1:200 for 2 hours at RT. Counterstain: DAPI (blue) nuclear stain at 1:40000 for 2 hours at RT. Localization: Cytoplasm. Localizes to the nucleus upon hypoxia. Magnification: 20x. (A) DAPI (blue) nuclear stain. (B) Anti-HIF2 alpha Antibody. (C) Composite. Product Citations (3) Western Blot Bp5250 inhibits vascular endothelial growth factor-induced angiogenesis and HIF-1α expression on endothelial cells. Lin, K., Lien J., Chung, C., Kuo, S. and Huang, T. (2012) Naunyn Schmiedebergs Arch Pharmacol. 385 (1): 39-49. PubMed ID: 21947252 Reactivity Human Applications: Western Blot Other Citations 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 MicroarrayBiotinProperties:Binds tetrameric avidin proteins including Streptavidin and neuravidin with very high affinityMolar mass: 244.31 g/molFormula: C10H16N2O3SApplications: Western blot, immunohistochemistry, and ELISABiotin Datasheet