Overview: Product Name Calnexin-NT Antibody DescriptionRabbit Anti-Human Calnexin-NT Polyclonal Species Reactivity Dog, Human, Monkey, Mouse, Rat, African clawed frog (Xenopus laevis), Bovine, Chicken, Guinea Pig (Cavia porcellus), Hamster, Pig, Rabbit, Sheep Applications WB, IHC, ICC/IF, IP Antibody Dilution WB (1:5000), IHC (1:100), ICC/IF (1:100), IP (1:100); optimal dilutions for assays should be determined by the user. Host Species Rabbit Immunogen Species Human Immunogen A 19 residue synthetic peptide based on dog calnexin and the peptide coupled to KLH 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 Rabbit Antiserum Storage Temperature -20ºC Shipping Temperature Blue Ice or 4ºC Purification Rabbit antiserum Clonality Polyclonal Specificity Detects the N-terminal domain of Calnexin ~90kDa. Cite This Product StressMarq Biosciences Cat# SPC-127, RRID: AB_2068995 Certificate of Analysis A 1:5000 dilution of SPC-127 was sufficient for detection of Calnexin in 20 µg of HeLa cell lysate by ECL immunoblot analysis. Biological Description Alternative Names Calnexin antibody, CALX_HUMAN antibody, CANX antibody, CNX antibody, FLJ26570 antibody, Histocompatibility complex class I antigen binding protein p88 antibody, IP90 antibody, Major histocompatibility complex class I antigen-binding protein p88 antibody, P90 antibody Research Areas Cell Signaling, Organelle Markers, Tags and Cell Markers Cellular Localization Endoplasmic Reticulum, Endoplasmic reticulum membrane, Melanosome Accession Number NP_001003232.1 Gene ID 403908 Swiss Prot P24643 Scientific Background Calnexin, an abundant ~90kDa integral protein of the endoplasmic reticulum, is also referred to as IP90, p88 and p90 (1). It consists of a large 50kDa N-terminal calcium-binding luminal domain, a single transmembrane helix and a short acidic cytoplasmic tail (2, 3). Unlike its ER counterparts which have a KDEL sequence on their C-terminus to ensure ER retention (4), calnexin has positively charged cytosolic residues that do the same thing (3). Most ER proteins act as molecular chaperones and participate in the proper folding of polypeptides and their assembly into mulit-subunit proteins. Calnexin together with calreticulin, plays a key role in glycoprotein folding and its control within the ER, by interacting with folding intermediates via their mono-glycosylated glycans (5, 6). Calnexin has also been shown to associate with the major histocompatibility complex class I heavy chains, partial complexes of the T cell receptor and B cell membrane immunoglobulin (7). References 1. Rajagopalan S., Xu Y., and Brenner M.B. (1994) Science 263(5145): 387-90. 2. Tjoelker L.W., et al. (1994) Biochemistry 33:3229. 3. Schrag J. et al. (2001) Molecular Cell 8(3): 633-644. 4. Janiszewski M. (2005) J. Biol Chem. 280(49):40813-40819. 5. Elagoz A., Callejo M., Armstrong J., and Rokeach L. A. (1999) J. Cell Sci. 112: 4449-4460. 6. Otteken A. and Moss B. (1996) J Bio Chem. 271(1): 97-103. 7. Galvin K. et al. (1992) Proc Natl Acad Sci USA. 89(18): 8452-6. 8. Raggo C., et al. (2002) Mol Cell Biol. 22: 5639-5649. 9. Rubio M.E., and Wenthold R.J. (1999) J Neurochem. 73: 942-948. Product Images Immunocytochemistry/Immunofluorescence analysis using Rabbit Anti-Calnexin Polyclonal Antibody (SPC-127). Tissue: HeLa cells. Species: Human. Primary Antibody: Rabbit Anti-Calnexin Polyclonal Antibody (SPC-127) at 1:100. Secondary Antibody: FITC Goat Anti-Rabbit (green). Western blot analysis of Human HeLa cell lysates showing detection of Calnexin protein using Rabbit Anti-Calnexin Polyclonal Antibody (SPC-127). Primary Antibody: Rabbit Anti-Calnexin Polyclonal Antibody (SPC-127) at 1:1000. Immunohistochemistry analysis using Rabbit Anti-Calnexin Polyclonal Antibody (SPC-127). Tissue: backskin. Species: Mouse. Fixation: Bouin’s Fixative Solution. Primary Antibody: Rabbit Anti-Calnexin Polyclonal Antibody (SPC-127) at 1:100 for 1 hour at RT. Secondary Antibody: FITC Goat Anti-Rabbit (green) at 1:50 for 1 hour at RT. Localization: Upper layer staining and few basal cell cytoplasmic staining. Western blot analysis of Rat tissue mix showing detection of Calnexin protein using Rabbit Anti-Calnexin Polyclonal Antibody (SPC-127). Load: 15 µg protein. Block: 1.5% BSA for 30 minutes at RT. Primary Antibody: Rabbit Anti-Calnexin Polyclonal Antibody (SPC-127) at 1:5000 for 2 hours at RT. Secondary Antibody: Donkey Anti-Rabbit IgG: HRP for 1 hour at RT. Immunohistochemistry analysis using Rabbit Anti-Calnexin Polyclonal Antibody (SPC-127). Tissue: colon colitis. Species: Mouse. Fixation: Formalin. Primary Antibody: Rabbit Anti-Calnexin Polyclonal Antibody (SPC-127) at 1:100000 for 12 hours at 4°C. Secondary Antibody: Biotin Goat Anti-Rabbit at 1:2000 for 1 hour at RT. Counterstain: Methyl Green at 200uL for 2 min at RT. Localization: Inflammatory cells. Product Citations (1) Immunocytochemistry/Immunofluorescence β1D chain increases α7β1 integrin and laminin and protects against sarcolemmal damage in mdx mice. Liu, J., Milner, D.J., Boppart, M.D., Ross, R.S. and Kaufman, S.J. (2012) Hum Mol Genet. 21 (7): 1592-1603. PubMed ID: 22180459 Reactivity Mouse Applications: Immunocytochemistry/Immunofluorescence  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