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ENTPD5 anticorps (Center)

ENTPD5 Reactivité: Humain WB, IHC (p) Hôte: Lapin Polyclonal unconjugated
N° du produit ABIN2856876
  • Antigène Voir toutes ENTPD5 Anticorps
    ENTPD5 (Ectonucleoside Triphosphate diphosphohydrolase 5 (ENTPD5))
    Épitope
    • 15
    • 7
    • 7
    • 6
    • 3
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    Center
    Reactivité
    • 49
    • 13
    • 5
    • 2
    • 2
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    Humain
    Hôte
    • 46
    • 3
    • 1
    Lapin
    Clonalité
    • 47
    • 3
    Polyclonal
    Conjugué
    • 19
    • 4
    • 3
    • 3
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    Cet anticorp ENTPD5 est non-conjugé
    Application
    • 41
    • 21
    • 13
    • 13
    • 5
    • 3
    • 3
    • 3
    • 2
    • 1
    Western Blotting (WB), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p))
    Attributs du produit
    Rabbit polyclonal antibody to ENTPD5(CD39L4) (ectonucleoside triphosphate diphosphohydrolase 5)
    ENTPD5 antibody [N1N3]
    Purification
    Purified by antigen-affinity chromatography.
    Immunogène
    Recombinant protein encompassing a sequence within the center region of human ENTPD5.
  • Indications d'application
    WB: 1:500-1:3000, IHC-P: 1:100-1:1000.
    Not yet tested in other applications.
    Optimal working dilutions should be determined experimentally by the end user.
    Restrictions
    For Research Use only
  • Validation #101762 (Western Blotting)
    'Independent Validation' signe
    by
    Department of Biochemistry and Molecular Genetics, University of Illinois at Chicago
    No.
    #101762
    Date
    02.11.2017
    Antigène
    ENTPD5
    Numéro du lot
    40128
    Application validée
    Western Blotting
    Contrôle positif
    Tet-inducible ENTPD5 LNCaP cells, Tet-inducible ENTPD5 BPH1 cells
    Contrôle négative
    BPH1 cells
    Conclusion

    Passed. ABIN2856876 specifically recognizes ENTPD5 in Tet-inducible ENTPD5 LNCaP cells and Tet-inducible ENTPD5 BPH1 cells.

    'Independent Validation' signe
    Validation Images
    Protocole
    Anticorps primaire
    ABIN2856876
    Anticorps secondaire
    HRP-conjugated goat-anti rabbit antibody (ThermoFisher, A16104, lot 42-28-042114)
    Full Protocol
    • Grow
      • LNCaP cells in RPMI 1640 1x with L-glutamine medium (Corning Cellgro, 10-040-CV, lot 11717002) supplemented with 10% FBS (GE Healthcare, SH30910.03, lot AC10225544) and penicillin-streptomycin (Mediatech, MT30002CI, lot 30002292) at 37°C and 5% CO2 in 5ml on a tissue culture dish to 1.5x106 cells/ml.
      • Tet-inducible GFP-LNCaP cells in RPMI 1640 1x with L-glutamine medium supplemented with 10% FBS, penicillin-streptomycin, and 1µg/ml Doxycycline hyclate (Sigma Aldrich, D9891, 056M4029V) at 37°C and 5% CO2 in 5ml on a tissue culture dish to 1.5x106 cells/ml.
      • Tet-inducible ENTPD5-LNCaP cells in RPMI 1640 1x with L-glutamine medium and 1µg/mL Doxycycline hyclate at 37°C and 5% CO2 in 5ml on a tissue culture dish to 1.5x106 cells/ml.
      • BPH1 cells in RPMI 1640 1x with L-glutamine medium supplemented with 10% FBS and penicillin-streptomycin at 37°C and 5% CO2 in 5ml on a tissue culture dish to 1.5x106 cells/ml.
      • Tet-inducible ENTPD5-BPH1 cells in RPMI 1640 1x with L-glutamine medium and 1µg/ml Doxycycline hyclate (Sigma Aldrich, D9891) at 37°C and 5% CO2 in 5ml on a tissue culture dish to 1.5x106 cells/ml.
    • Collect and resuspend cells in 50µl lysis buffer (20mM HEPES pH7.4, 1% Triton X-100, 150mM NaCl, 1mM EGTA, 1mM EDTA, 10mM sodium pyrophosphate, 100mM NaF, 5mM iodoacetic acid, 20nM okadeic acid, 0.2mM PMSF, and 1 tablet/10mL Pierce Protease Inhibitor mini Tablet (ThermoFisher, 88665, Lot RC2161121)).
    • Lyse cells by 5 rounds of freeze/thaw cycles: 60sec in LN2, 30sec in a 37°C water bath, 15sec vortex, 60sec on ice.
    • Spin lysed cells at 10000xg for 5min at 4°C.
    • Recover the supernatant and measure protein concentration with Bradford reagent (Bio-Rad, 5000006).
    • Denature 25μg total proteins of each sample for 5min at 95°C in Laemmli SDS sample buffer and subsequently separate them on two separate 4-12% Express Plus Page Gels (GenScript, M41215, Lot:C30251704) for 40min at 160V in an electrophoresis chamber (BioRad).
    • Transfer proteins onto PVDF membrane (Millipore, IPFL00010, Lot K0MA0558B) with a wet blotting system (Bio-Rad) for 75min at 115V at 4°C.
    • Block the membrane with 5% non-fat dry milk in TBSTween (0.1% v/v) for 1h at RT.
    • Incubation the membranes with primary
      • rabbit anti-ENTPD5 antibody (antibodies-online, ABIN2856876, Lot: 40128) or
      • rabbit anti-ENTPD5 antibody (Sigma, HPA002927 Lot: A40077)
    • diluted 1:500 in 5% non-fat dry milk in TBST at for 16h at 4°C.
    • Wash the membrane 5x for 5min in TBST.
    • Incubate the membrane with secondary HRP-conjugated goat-anti rabbit antibody (ThermoFisher, A16104, lot 42-28-042114) diluted 1:3000 in 5% non-fat dry milk in TBST for 1h at RT.
    • Wash membrane 5x for 5min with PBST.
    • Visualize protein bands using SuperSignal West Pico Chemiluminescent Substrate (ThermoScientific, 34087, Lot SC246874) on CL-X Posure Film (ThermoScientific, 34091).
    Notes
    • The ENTPD5 antibody ABIN2856876 detects a single band around expected molecular weight (47kDa). The intensity of this band becomes more intense upon ENTPD5 overexpression and disappears in the negative control cell line.

    • ABIN2856876 also reveals fewer non-specific bands than the control antibody. We recommend this antibody for detecting ENTPD5. The positive control antibody performed as predicted with showing increased expression in the overexpression cells and no expression in the negative control BPH1 cell line.

  • Format
    Liquid
    Concentration
    1 mg/mL
    Buffer
    0.1M Tris, 0.1M Glycine, 10 % Glycerol ( pH 7).
    0.01 % Thimerosal was added as a preservative.
    Agent conservateur
    Thimerosal (Merthiolate)
    Précaution d'utilisation
    This product contains Thimerosal (Merthiolate): a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
    Stock
    -20 °C
    Stockage commentaire
    Keep as concentrated solution. Aliquot and store at -20°C or below.
    Avoid multiple freeze-thaw cycles.
  • Antigène
    ENTPD5 (Ectonucleoside Triphosphate diphosphohydrolase 5 (ENTPD5))
    Autre désignation
    ENTPD5 (ENTPD5 Produits)
    Synonymes
    anticorps ENTPD5, anticorps CD39L4, anticorps NTPDase-5, anticorps PCPH, anticorps AI196558, anticorps AI987697, anticorps Cd39l4, anticorps ER-UDPase, anticorps NTPDase5, anticorps Pcph, anticorps mNTPase, anticorps cph, anticorps ectonucleoside triphosphate diphosphohydrolase 5, anticorps ectonucleoside triphosphate diphosphohydrolase 5 L homeolog, anticorps ENTPD5, anticorps entpd5, anticorps entpd5.L, anticorps Entpd5
    ID gène
    957
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