Telefon:
+49 (0)241 95 163 153
Fax:
+49 (0)241 95 163 155
E-Mail:
orders@antikoerper-online.de

ENTPD5 Antikörper (Center)

ENTPD5 Reaktivität: Human WB, IHC (p) Wirt: Kaninchen Polyclonal unconjugated
Produktnummer ABIN2856876
  • Target Alle ENTPD5 Antikörper anzeigen
    ENTPD5 (Ectonucleoside Triphosphate diphosphohydrolase 5 (ENTPD5))
    Bindungsspezifität
    • 15
    • 7
    • 7
    • 6
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    Center
    Reaktivität
    • 50
    • 13
    • 5
    • 2
    • 2
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    Human
    Wirt
    • 47
    • 3
    • 1
    Kaninchen
    Klonalität
    • 48
    • 3
    Polyklonal
    Konjugat
    • 20
    • 4
    • 3
    • 3
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    Dieser ENTPD5 Antikörper ist unkonjugiert
    Applikation
    • 42
    • 21
    • 13
    • 13
    • 5
    • 3
    • 3
    • 3
    • 2
    • 1
    Western Blotting (WB), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p))
    Produktmerkmale
    Rabbit polyclonal antibody to ENTPD5(CD39L4) (ectonucleoside triphosphate diphosphohydrolase 5)
    ENTPD5 antibody [N1N3]
    Aufreinigung
    Purified by antigen-affinity chromatography.
    Immunogen
    Recombinant protein encompassing a sequence within the center region of human ENTPD5.
  • Applikationshinweise
    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.
    Beschränkungen
    Nur für Forschungszwecke einsetzbar
  • Validierung #101762 (Western Blotting)
    'Independent Validation' Siegel
    by
    Department of Biochemistry and Molecular Genetics, University of Illinois at Chicago
    No.
    #101762
    Datum
    02.11.2017
    Antigen
    ENTPD5
    Chargennummer
    40128
    Validierte Anwendung
    Western Blotting
    Positivkontrolle
    Tet-inducible ENTPD5 LNCaP cells, Tet-inducible ENTPD5 BPH1 cells
    Negativkontrolle
    BPH1 cells
    Bewertung

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

    'Independent Validation' Siegel
    Validierungsbilder
    Protokoll
    Primärantikörper
    ABIN2856876
    Sekundärantikörper
    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).
    Anmerkungen
    • 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
    Konzentration
    1 mg/mL
    Buffer
    0.1M Tris, 0.1M Glycine, 10 % Glycerol ( pH 7).
    0.01 % Thimerosal was added as a preservative.
    Konservierungsmittel
    Thimerosal (Merthiolate)
    Vorsichtsmaßnahmen
    This product contains Thimerosal (Merthiolate): a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
    Lagerung
    -20 °C
    Informationen zur Lagerung
    Keep as concentrated solution. Aliquot and store at -20°C or below.
    Avoid multiple freeze-thaw cycles.
  • Target
    ENTPD5 (Ectonucleoside Triphosphate diphosphohydrolase 5 (ENTPD5))
    Andere Bezeichnung
    ENTPD5 (ENTPD5 Produkte)
    Synonyme
    ENTPD5 antikoerper, CD39L4 antikoerper, NTPDase-5 antikoerper, PCPH antikoerper, AI196558 antikoerper, AI987697 antikoerper, Cd39l4 antikoerper, ER-UDPase antikoerper, NTPDase5 antikoerper, Pcph antikoerper, mNTPase antikoerper, cph antikoerper, ectonucleoside triphosphate diphosphohydrolase 5 antikoerper, ectonucleoside triphosphate diphosphohydrolase 5 L homeolog antikoerper, ENTPD5 antikoerper, entpd5 antikoerper, entpd5.L antikoerper, Entpd5 antikoerper
    Gen-ID
    957
Sie sind hier:
Kundenservice