Overview: Product Name ATG12 Antibody DescriptionRabbit Anti-Human ATG12 Polyclonal 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 Rabbit Immunogen Species Human Immunogen Synthetic peptide from the C-terminal of human ATG12 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, 50% glycerol, 0.09% sodium azide Storage Temperature -20ºC Shipping Temperature Blue Ice or 4ºC Purification Peptide Affinity Purified Clonality Polyclonal Specificity Predicted molecular weight at ~15.1kDa. Observed molecular weights at ~48-55kDa based on ATG12-ATG5 heterodimer. Cite This Product StressMarq Biosciences Cat# SPC-608, RRID: AB_2704853 Certificate of Analysis A 1:1000 dilution of SPC-608 was sufficient for detection of ATG12 on HeLa cell lysates using Goat anti-rabbit IgG:HRP as the secondary antibody. Biological Description Alternative Names APG12-like Antibody, APG12 Antibody, ATG12_Human Antibody, HAPG12 Antibody, FBR93 Antibody Research Areas Cancer, Autophagy, Cardiovascular System, Cell Signaling, Epigenetics and Nuclear Signaling, Heart Cellular Localization Cytoplasm Accession Number NP_001264712.1 Gene ID 9140 Swiss Prot O94817 Scientific Background ATG12 (autophagy-related protein 12), also known as APG12, is a 140 amino acid protein that is ubiquitously expressed and belongs to the ATG12 family of proteins. ATG12 is a homolog of the yeast protein APG12 that participates in autophagy. Autophagy is a membrane trafficking mechanism that delivers cytoplasmic cargo to the vacuole/lysosome for degradation and recycling. In yeast, autophagy requires a protein conjugation system consisting of APG12 covalently bound at the carboxy terminal glycine to lysine 149 of APG5. Similarly in humans, ATG12 is essential for autophagy and localizes to the cytoplasm where it is covalently bound to APG5, a conjugation reaction that requires APG7, ATG10 and ATP (1-3). The ATG12-APG5 conjugate functions as an important regulator of the autophagic process and is required for the change in membrane morphology and development of autophagosomes. Due to alternative splicing events, two ATG12 isoforms exist. References 1. Mizushima N., Sugita H., Yoshimori T., Ohsumi Y. (1999) J Biol Chem. 273(51): 33889-33892. 2. Mizushuma N., et al. (1998) Nature. 395: 395-398. 3. Suzuki K., et al. (2001) EMBO J. 20: 5971-5981. Product Images Immunocytochemistry/Immunofluorescence analysis using Rabbit Anti-ATG12 Polyclonal Antibody (SPC-608). Tissue: Cervical Cancer cell line (HeLa). Species: Human. Fixation: 4% Formaldehyde for 15 min at RT. Primary Antibody: Rabbit Anti-ATG12 Polyclonal Antibody (SPC-608) at 1:100 for 60 min at RT. Secondary Antibody: Goat Anti-Rabbit ATTO 488 at 1:100 for 60 min at RT. Counterstain: DAPI (blue) nuclear stain at 1:5000 for 5 min RT. Localization: Nucleus, Cytoplasm. Magnification: 40X. (A) DAPI (blue) nuclear stain (B) Phalloidin Texas Red F-Actin stain (C) ATG12 Antibody (D) Composite. Western blot analysis of Human HeLa cell lysates showing detection of ~15kDa ATG12 protein using Rabbit Anti-ATG12 Polyclonal Antibody (SPC-608). Lane 1: MW Ladder. Lane 2: Human HeLa (20 µg). Load: 20 µg. Block: 5% milk + TBST for 1 hour at RT. Primary Antibody: Rabbit Anti-ATG12 Polyclonal Antibody (SPC-608) at 1:1000 for 1 hour at RT. Secondary Antibody: Goat Anti-Rabbit: HRP at 1:2000 for 1 hour at RT. Color Development: TMB solution for 12 min at RT. Predicted/Observed Size: ~15kDa. Other Band(s): ATG12-ATG5 Complex. Product Citations (0) Currently there are no citations for this product.  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