Overview: Product Name Calreticulin Antibody DescriptionRabbit Anti-Human Calreticulin Polyclonal Species Reactivity Dog, Human, Monkey, Mouse, Rat, Bovine, Chicken, Guinea Pig (Cavia porcellus), Hamster, Pig, Rabbit, Sheep Applications WB, IHC, ICC/IF, IP, FCM Antibody Dilution WB (1:1000), IHC (1:100), ICC/IF (1:100); optimal dilutions for assays should be determined by the user. Host Species Rabbit Immunogen Species Human Immunogen Human calreticulin synthetic peptide with a cysteine residue added and the peptide conjugated to KLH 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 Peptide Affinity Purified Clonality Polyclonal Specificity Detects ~63kDa. Cite This Product StressMarq Biosciences Cat# SPC-122, RRID: AB_2069601 Certificate of Analysis A 1:1000 dilution of SPC-122 was sufficient for detection of Calreticulin in 20 µg of HeLa cell lysate by ECL immunoblot analysis. Biological Description Alternative Names CALR Antibody, Calregulin Antibody, cC1qR Antibody, CRP55 Antibody, ERp60 Antibody, HSCBP Antibody, RO Antibody, SSA Antibody, grp60 Antibody Research Areas Cell Signaling, Protein Trafficking Cellular Localization Cytoplasm, Cell Surface, Endoplasmic Reticulum, Endoplasmic reticulum lumen, Extracellular Matrix, Sarcoplasmic Reticulum, Sarcoplasmic Reticulum Lumen Accession Number NP_004334.1 Gene ID 811 Swiss Prot P27797 Scientific Background Calreticulin is a multifunctional, highly conserved Ca2+ -binding protein that is localized to the endoplasmic reticulum (ER), but has also been detected in the nucleus and nuclear envelop. Like many other ER proteins, it has the conserved ER retention KDEL (Lys-Asp-Glu-Leu) sequence at its C-terminus (1-3). CRT’s three domains include a 180 residue N-terminal domain, a proline-rich P-domain (residues 189-288) that binds Ca2+ with high affinity and shares homology with calnexin (CNX) and calmegin, and a 110 residue C-terminal domain that binds Ca2+ with low affinity but high capacity (1,3). Recent studies suggest that this soluble ER protein has a multifunctional role. It appears to be involved in calcium storage and regulation as well as having a molecular chaperone activity. It has been shown to interact with the cytoskeleton and to be involved in the regulation of gene expression. Calreticulin may also play a role in cellular proliferation including its apparent activity in the proliferation of certain viruses within mammalian host cells (4, 5), and it has also been shown to be induced in response to various types of cell stress including amino acid deprivation (6). Close interconnections among protein synthesis, gene expression and calcium signaling have been observed by many researchers in recent years. Calreticulin might be centrally located and therefore it crucially participates in the coordination of many functions by the cell (4, 5). Studies also suggest its involvement in a few diseases such as systemic lupus erythematosus, rheumatoid arthritis, celiac disease, complete congenital heart block, and halothane hepatitis (1). References 1. Johnson S., et al. (2001) Trends Cell Biol 11: 122-129. 2. Smith M.J., et al. (1989) EMBO J. 8: 3581-3586. 3. Ellgaard L., et al. (2001) Curr Opin Cell Biol. 13: 431-437. 4. Krause K.H., and Michalak M. (1997) Cell. 88: 439-443. 5. Nash P.D., et al. (1994) Mol Cell Biochem. 135: 71-78. 6. Heal R., and McGivan J. (1998) Biochem J. 329: 389-394. 7. Lucero H.A., et al. (1998) J Biol Chem. 273: 9857-9863. 8. Tanaka S., et al. (2000) J Biol Chem. 275: 10388-10393. 9. Yoon G.S., et al. (2000) Cancer Res. 60: 1117-1120. 10. Antoniou A.N., et al. (2002) Immunology 106: 182-189. 11. Wada I., et al. (1997) EMBO J. 16: 5420-5432. 12. Laguerre D.B., et al. (1998) J Vir. 72: 4940-4949. Product Images Immunocytochemistry/Immunofluorescence analysis using Rabbit Anti-Calreticulin Polyclonal Antibody (SPC-122). Tissue: Heat Shocked HeLa Cells. Species: Human. Fixation: 2% Formaldehyde for 20 min at RT. Primary Antibody: Rabbit Anti-Calreticulin Polyclonal Antibody (SPC-122) at 1:100 for 12 hours at 4°C. Secondary Antibody: FITC Goat Anti-Rabbit (green) at 1:200 for 2 hours at RT. Counterstain: DAPI (blue) nuclear stain at 1:40000 for 2 hours at RT. Localization: Endoplasmic reticulum lumen. Cytoplasm. Magnification: 100x. (A) DAPI (blue) nuclear stain. (B) Anti-Calreticulin Antibody. (C) Composite. Heat Shocked at 42°C for 1h. Western blot analysis of multiple cell lines lysates showing detection of Calreticulin protein using Rabbit Anti-Calreticulin Polyclonal Antibody (SPC-122). Load: 15 µg protein. Block: 1.5% BSA for 30 minutes at RT. Primary Antibody: Rabbit Anti-Calreticulin Polyclonal Antibody (SPC-122) 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-Calreticulin Polyclonal Antibody (SPC-122). Tissue: backskin. Species: Mouse. Fixation: Bouin’s Fixative Solution. Primary Antibody: Rabbit Anti-Calreticulin Polyclonal Antibody (SPC-122) at 1:100 for 1 hour at RT. Secondary Antibody: FITC Goat Anti-Rabbit (green) at 1:50 for 1 hour at RT. Localization: Cytoplasmic granule. Endoplasmic reticulum lumen. Immunocytochemistry/Immunofluorescence analysis using Rabbit Anti-Calreticulin Polyclonal Antibody (SPC-122). Tissue: Heat Shocked HeLa Cells. Species: Human. Fixation: 2% Formaldehyde for 20 min at RT. Primary Antibody: Rabbit Anti-Calreticulin Polyclonal Antibody (SPC-122) at 1:100 for 12 hours at 4°C. Secondary Antibody: R-PE Goat Anti-Rabbit (yellow) at 1:200 for 2 hours at RT. Counterstain: DAPI (blue) nuclear stain at 1:40000 for 2 hours at RT. Localization: Endoplasmic reticulum lumen. Cytoplasm. Magnification: 20x. (A) DAPI (blue) nuclear stain. (B) Anti-Calreticulin Antibody. (C) Composite. Heat Shocked at 42°C for 1h. Product Citations (7) Western Blot Inhibition of receptor activator of nuclear factor-κB ligand- or lipopolysaccharide-induced osteoclast formation by conophylline through downregulation of CREB. Koide, N. et al. (2014) Immunol Lett. 161(1):31-7.  PubMed ID: 24792671 Reactivity Mouse Applications: Western Blot Ascorbic acid deficiency decreases hepatic cytochrome P-450, especially CYP2B1/2B2, and simultaneously induces heme oxygenase-1 gene expression in scurvy-prone ODS rats. Kobayashi, M. et al. (2014) Biosci Biotechnol Biochem. 78(6):1060-6. PubMed ID: 25036135 Reactivity Rat Applications: Western Blot Immunogenic chemotherapy with cyclophosphamide and doxorubicin against established murine carcinoma. Tongu, M., Harashima, N., Yamada, T., Harada, T. and Harada, M. (2010) Cancer Immunol Immunother. 59 (5): 769-777. PubMed ID: 19940990 Reactivity Mouse Applications: Western Blot Immunohistochemistry Calreticulin is a novel independent prognostic factor for oral squamous cell carcinoma. Harada, K. et al. (2017) Oncol lett. 13(6): 4857-4862. PubMed ID: N/A Reactivity Human Applications: Immunohistochemistry Flow Cytometry Independent of Plasmacytoid Dendritic Cell (pDC) infection, pDC Triggered by Virus-Infected Cells Mount Enhanced Type I IFN Responses of Different Composition as Opposed to pDC Stimulated with Free Virus. Frenz, T. et al. (2014) J Immunol. 193(5):2496-503. PubMed ID: 25070849 Reactivity Mouse Applications: Flow Cytometry Immunocytochemistry/Immunofluorescence HIV-1 Vpu Antagonizes CD317/tetherin by Adaptor protein-1-Mediated Exclusion from Virus Assembly Sites. Pujol, F.M. et al. (2016) J Virol. 90(15):6709-23. PubMed ID: 27170757 Reactivity Human Applications: Immunocytochemistry/Immunofluorescence Nef Proteins of Epidemic HIV-1 Group O Strains Antagonize Human Tetherin. Kluge, S.F. et al. (2014) Cell Host Microbe. 16(5):639-50. PubMed ID: 25525794 Reactivity Human Applications: Immunocytochemistry/ImmunofluorescenceAPC (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