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GST-Tag antibody

Reactivity: Schistosoma japonicum WB, IP, IHC, ChIP Host: Mouse Monoclonal 5C12 unconjugated
Catalog No. ABIN3045984
  • Target See all GST-Tag products
    GST-Tag
    Reactivity
    • 32
    • 6
    • 2
    Schistosoma japonicum
    Host
    • 23
    • 17
    • 3
    • 1
    • 1
    Mouse
    Clonality
    • 26
    • 19
    Monoclonal
    Conjugate
    • 27
    • 4
    • 3
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    This GST-Tag antibody is un-conjugated
    Application
    • 36
    • 15
    • 10
    • 4
    • 1
    • 1
    • 1
    • 1
    Western Blotting (WB), Immunoprecipitation (IP), Immunohistochemistry (IHC), Chromatin Immunoprecipitation (ChIP)
    Characteristics
    The GST tag issued in many protocols
    Purification
    purified form hybridoma supernatant
    Immunogen
    GST
    Clone
    5C12
    Isotype
    IgG1
  • Application Notes
    WB 0.1-1.0ug/ml
    Restrictions
    For Research Use only
  • Validation #029860 (Western Blotting)
    'Independent Validation' Badge
    by
    Celplor LLC
    No.
    #029860
    Date
    05/19/2016
    Antigen
    Mouse anti-GST monoclonal antibody
    Lot Number
    1214
    Method validated
    Western Blotting
    Positive Control
    BL21 bacteria cells were transformed with an in-house GST expression vector pGST
    Negative Control
    Empty vector cell lysate
    Notes
    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' Badge
    Validation Images
    Full Methods
    Primary Antibody
    • Antibody: Mouse anti-GST monoclonal antibody
    • Lot number: 1214
    • Dilution: 1:1000
    Secondary Antibody
    • 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).
    Experimental Notes
  • Format
    Liquid
    Concentration
    1 mg/mL
    Buffer
    PBS pH7.2
    Storage
    -20 °C
  • Target
    GST-Tag
    Alternative Name
    GST Tag (GST-Tag Products)
    Target Type
    Tag
    Molecular Weight
    26 kDa
    UniProt
    P08515
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