GST-Tag Antikörper

Details zu Produkt Nr. ABIN3045984, Anbieter: Anmelden zum Anzeigen
Antigen
Reaktivität
Schistosoma japonicum
78
56
26
14
8
3
1
1
1
Wirt
Maus
121
24
14
4
1
Klonalität (Klon)
Monoklonal ()
Konjugat
Unkonjugiert
11
7
7
6
6
6
6
5
5
5
5
5
5
5
5
5
5
5
2
1
1
1
1
1
Applikation
Chromatin Immunoprecipitation (ChIP), Immunoprecipitation (IP), Immunohistochemistry (IHC), Western Blotting (WB)
152
81
23
20
17
10
6
2
1
1
1
Optionen
Hersteller
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Hersteller Produkt- Nr.
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'Independent Validation' Siegel
Antigen Mouse anti-GST monoclonal antibody
Chargennummer 1214
Validierte Anwendung Western Blotting
Positivkontrolle BL21 bacteria cells were transformed with an in-house GST expression vector pGST
Negativkontrolle Empty vector cell lysate
Bewertung 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.
Primärantikörper
Sekundärantikörper
Kontrollen
  • Positive control: BL21 bacteria cells were transformed with an in-house GST expression vector pGST
  • Negative control: Empty vector cell lysate
Protokoll
  • **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).
Validierungsbilder
Western Blotting validation image for anti-GST-Tag antibody (ABIN3045984) Figure 1: GST content of BL21-pGST(+IPTG) is 12% of total protein. GST content in IPT...
Western Blotting validation image for anti-GST-Tag antibody (ABIN3045984) Figure 2: Western blot for mouse anti-GST monoclonal antibody. The first lane shows t...
Immunogen GST
Klon 5C12
Isotyp IgG1
Produktmerkmale The GST tag issued in many protocols
Reinigung purified form hybridoma supernatant
Andere Bezeichnung GST Tag
Molekulargewicht 26 kDa
UniProt P08515
Applikationshinweise WB 0.1-1.0ug/ml
Beschränkungen Nur für Forschungszwecke einsetzbar
Format Liquid
Konzentration 1 mg/mL
Buffer PBS pH7.2
Lagerung -20 °C
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