Overview: Product Name CD74 Antibody DescriptionMouse Anti-Human CD74 Monoclonal IgG2a Species Reactivity Human Applications WB, ICC/IF Antibody Dilution WB (1:1000); ICC/IF (1:100); optimal dilutions for assays should be determined by the user. Host Species Mouse Immunogen Species Human Immunogen Human CD74 invariant chain synthetic peptide. 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 pH 7.4, 50% glycerol, 0.09% Sodium Azide Storage Temperature -20ºC Shipping Temperature Blue Ice or 4ºC Purification Protein G Purified Clonality Monoclonal Clone Number 1B8 Isotype IgG2a Specificity Detects ~33 kDa protein. Cite This Product StressMarq Biosciences Cat# SMC-267, RRID: AB_2699952 Certificate of Analysis A 1:1000 dilution of SMC-267 was sufficient for detection of CD74 in 15 µg of B-Cell Lymphoma cell line Raji by ECL immunoblot analysis using Goat Anti-Mouse IgG:HRP as the secondary Antibody. Biological Description Alternative Names DHLAG Antibody, HLA DR gamma Antibody, HLADG Antibody, p33 Antibody, p35 Antibody, Protein 41 Antibody, CD 74 Antibody, CD74 Antibody, CD74 antigen (invariant polypeptide of major histocompatibility complex, class II antigen-associated) Antibody, CD74 antigen Antibody, CD74 molecule Antibody, CD74 molecule, major histocompatibility complex, class II invariant chain Antibody, CLIP Antibody, DHLAG Antibody, Gamma chain of class II antigens Antibody, HG2A_HUMAN Antibody, HLA class II histocompatibility antigen gamma chain Antibody, HLA DR antigens associated invariant chain Antibody, HLA DR gamma Antibody, HLA-DR antigens-associated invariant chain Antibody, HLA-DR-gamma Antibody, HLADG Antibody, HLADR antigens associated invariant chain Antibody Ia antigen associated invariant chain Antibody, Ia antigen-associated invariant chain Antibody, Ia associated invariant chain Antibody, Ia gamma Antibody, Ii Antibody, Invariant polypeptide of major histocompatibility complex class II antigen associated Antibody, la-gamma Antibody, Major histocompatibility complex class II invariant chain Antibody, MHC HLA DR gamma chain Antibody, MHC HLA-DR gamma chain Antibody, p33 Antibody, p35 Antibody, Protein 41 Antibody Research Areas Adaptive Immunity, B Cell CD Markers, B Cells, Immunology Cellular Localization Cell membrane, Endoplasmic Reticulum, Endoplasmic reticulum membrane, Endosome, Golgi apparatus, Lysosome Accession Number NP_001020329.1 Gene ID 972 Swiss Prot P04233 Scientific Background CD74 is a non-polymorphic type II integral membrane protein. It has a short N-terminal cytoplasmic tail of 28 amino acids, followed by a single 24-aa transmembrane region and an approximately 150-aa lumenal domain (1). The CD74 chain is thought to function mainly as an MHC class II chaperone, which promotes ER exit of MHC class II molecules, directs them to endocytic compartments, prevents peptide binding in the ER, and contributes to peptide editing in the MHC class II compartment. Class II MHC and Ii expression was believed to be restricted to classical antigen-presenting cells (APC); however, during inflammation, other cell types, including mucosal epithelial cells, have also been reported to express class II MHC molecules (2). Experiments that investigate cell-surface CD74 are complicated by the fact that CD74 remains on the cell surface for a very short time. The surface half-life of CD74 was calculated to be fewer than 10 minutes (3). CD74 however has also recently been shown to have a role as an accessory-signaling molecule because of its high-affinity binding to the pro-inflammatory cytokine, macrophage migration-inhibitory factor (MIF) (3). The restricted expression of CD74 by normal tissues and its very rapid internalization make CD74 an attractive therapeutic target for both cancer and immunologic diseases (4). References 1. Becker-Hermann, S., Arie, G., Medvedovsky H, Kerem A, and Shachar I. (2005) Mol Bio Cell. 16(11):5061-9. 2. Barrera CA., et al (2005) J Histochem Cytochem 53 (12): 1481-9. 3. Starlets D., et al. (2006) Blood. 107 (12): 4807-4816. 4. Burton JD., et al. (2004). Clin Cancer Res. 10(19): 6606-11. 5. Denzin L.K., Hammond, C. and Cresswell, P. (1996) J. Exp. Med. 184: 2153-2165. 6. Denzin L.K., Robbins N.F., Carboy-Newcomb C. and Cresswell P. (1994) Immunity 1: 595-606. Product Images Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-CD74 Monoclonal Antibody, Clone 1B8 (SMC-267). Tissue: HeLa Cells (Human Cervical Cancer). Species: Human. Fixation: 4% Formaldehyde for 15 min at RT. Primary Antibody: Mouse Anti-CD74 Monoclonal Antibody (SMC-267) at 1:100 for 60 min at RT. Secondary Antibody: Goat Anti-Mouse ATTO 488 at 1:200 for 60 min at RT. Counterstain: Phalloidin Texas Red F-Actin stain; DAPI (blue) nuclear stain at 1:1000, 1:5000 for 60 min at RT, 5 min at RT. Localization: Cell membrane, Endoplasmic Reticulum, Golgi apparatus. Magnification: 60X. (A) DAPI (blue) nuclear stain (B) Phalloidin Texas Red F-Actin stain (C) CD74 Antibody (D) Composite. Western Blot analysis of Human Lymphoblastoid cell line (Raji) showing detection of 33-35 kDa CD74 protein using Mouse Anti-CD74 Monoclonal Antibody, Clone 1B8 (SMC-267). Lane 1: Molecular Weight Ladder (MW). Lane 2: Raji cell lysate. Load: 15 µg. Block: 5% Skim Milk in TBST. Primary Antibody: Mouse Anti-CD74 Monoclonal Antibody (SMC-267) at 1:1000 for 2 hours at RT. Secondary Antibody: Goat Anti-Mouse IgG: HRP at 1:1000 for 60 min at RT. Color Development: ECL solution for 5 min in RT. Predicted/Observed Size: 33-35 kDa. Product Citations (0) Currently there are no citations for this product.BiotinProperties:Binds tetrameric avidin proteins including Streptavidin and neuravidin with very high affinityMolar mass: 244.31 g/molFormula: C10H16N2O3SApplications: Western blot, immunohistochemistry, and ELISABiotin Datasheet