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GST-Tag anticorps

Reactivité: Schistosoma japonicum WB, IP, IHC, ChIP Hôte: Souris Monoclonal 5C12 unconjugated
N° du produit ABIN3045984
  • Antigène Tous les produits GST-Tag
    GST-Tag
    Reactivité
    • 30
    • 6
    • 2
    Schistosoma japonicum
    Hôte
    • 23
    • 20
    • 3
    • 1
    • 1
    Souris
    Clonalité
    • 25
    • 23
    Monoclonal
    Conjugué
    • 30
    • 6
    • 3
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    Cet anticorp GST-Tag est non-conjugé
    Application
    • 41
    • 17
    • 11
    • 5
    • 1
    • 1
    • 1
    • 1
    • 1
    Western Blotting (WB), Immunoprecipitation (IP), Immunohistochemistry (IHC), Chromatin Immunoprecipitation (ChIP)
    Attributs du produit
    The GST tag issued in many protocols
    Purification
    purified form hybridoma supernatant
    Immunogène
    GST
    Clone
    5C12
    Isotype
    IgG1
  • Indications d'application
    WB 0.1-1.0ug/ml
    Restrictions
    For Research Use only
  • Validation #029860 (Western Blotting)
    'Independent Validation' signe
    by
    Celplor LLC
    No.
    #029860
    Date
    19.05.2016
    Antigène
    Mouse anti-GST monoclonal antibody
    Numéro du lot
    1214
    Application validée
    Western Blotting
    Contrôle positif
    BL21 bacteria cells were transformed with an in-house GST expression vector pGST
    Contrôle négative
    Empty vector cell lysate
    Conclusion
    Based on the 12% GST content in total cell lysate of BL21-pGST (+IPTG), there is an estimated 1 ng of GST protein in 0.01 ug of total protein with IPTG induction. Therefore, the detection limit of mouse anti-GST monoclonal antibody is lower than 1 ng of GST protein.
    'Independent Validation' signe
    Validation Images
    Protocole
    Anticorps primaire
    • Antibody: Mouse anti-GST monoclonal antibody
    • Lot number: 1214
    • Dilution: 1:1000
    Anticorps secondaire
    • Antibody: HRP labeled secondary anti-mouse antibody
    • Lot number: 390630
    • Dilution: 1:2000
    Full Protocol
    • **I. Reagent preparation**
    • 1. BL21 bacteria cells were transformed with an in-house GST expression vector pGST.
    • 2. Individual colony was inoculated in 2xYT medium and cultured at 37°C with shaking for 5 hrs followed by IPTG (1mM) induction for 2 hrs. Bacteria was grown to OD600=0.6-0.8 (5 hr) when IPTG was added.
    • 3. Cells were centrifuged at top speed and pellets were collected.
    • 4. Cell pellets were lysed in SDS-PAGE sample buffer.
    • 5. Protein concentration was assayed using DC protein assay kit (Bio-Rad).
    • 6. 50 ug of cell lysate was loaded and protein was separated by SDS-PAGE.
    • 7. Gel was stained with SimplyBlue Safe Stain (Invitrogen) at room temperature.
    • 8. GST content in IPTG induced samples was analyzed by Quantity One software (Bio-Rad).
    • **II. Western blot validation**
    • 1. Cell lysates from BL21 host cells, BL21-pGST transformed cells with or without IPTG induction were loaded and proteins were separated by SDS-PAGE followed by Western transfer to a nitrocellulose membrane.
    • 2. Membrane was blocked with 2% powder milk (Bio-Rad) for 1 hr at room temperature.
    • 3. Membrane was incubated with mouse anti-GST monoclonal antibody (1:1000 dilution) at 4°C overnight with shaking.
    • 4. Membrane was washed with TBST three times for 10 min.
    • 5. After washing, membrane was incubated with HRP labeled secondary anti-mouse antibody (1:2000 dilution, GE Healthcare) with shaking at room temperature for 1 hr.
    • 6. Membrane was washed with TBST three times for 10 min.
    • 7. After washing, membrane was incubated with SuperSignal West Pico substrate (Thermo Scientific) for 5 min at room temperature.
    • 8. Membrane was analyzed by ChemiDoc XRS gel documentation system (Bio-Rad).
    Notes
  • Format
    Liquid
    Concentration
    1 mg/mL
    Buffer
    PBS pH7.2
    Stock
    -20 °C
  • Antigène
    GST-Tag
    Autre désignation
    GST Tag (GST-Tag Produits)
    Classe de substances
    Tag
    Poids moléculaire
    26 kDa
    UniProt
    P08515
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