GST Antikörper (Glutathione S Transferase)

Details for Product anti-GST Antibody No. ABIN1998461, Anbieter: Anmelden zum Anzeigen
Antigen
Reaktivität
Schistosoma japonicum
280
85
48
30
26
9
4
3
2
2
1
1
1
Wirt
Maus
224
145
94
5
5
4
2
1
Klonalität (Klon)
Monoklonal ()
Konjugat
Dieser GST Antikörper ist unkonjugiert
34
31
27
14
8
6
6
4
4
3
3
3
3
2
2
2
2
2
2
2
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
1
Applikation
Immunoprecipitation (IP), Western Blotting (WB)
387
236
95
89
40
31
17
15
12
11
9
9
9
9
7
6
5
3
3
2
2
2
1
1
1
1
1
1
1
1
Optionen
Hersteller
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Hersteller Produkt- Nr.
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'Independent Validation' Siegel
Antigen Glutathione S Transferase (GST)
Chargennummer HD04FE1006
Validierte Anwendung Western Blotting
Positivkontrolle GST- AcNFkBp(C-terminal) 293-Lysate
Negativkontrolle Empty vector- 293 Lysate
Bewertung A strong specific band was observed in the positive control at the expected size (~82 kDa) that is not observed in the negative control.
Primärantikörper
Sekundärantikörper
Kontrollen
  • Positive control: GST- AcNFkBp(C-terminal) 293-Lysate
  • Negative control: Empty vector- 293 Lysate
Protokoll
  • 1. The cell extracts were heated at 95°C for 5 minutes in 1X SDS Sample Buffer containing 1% SDS and 1.25% β-mercaptoethanol.
  • 2. 15 μl of heated were loaded and resolved on 8-16% SDS-polyacrylamide gel.
  • 3. The Bio-Rad Precision Plus (Cat 161-0374) were used as molecular mass markers.
  • 4. Proteins were then transferred onto PVDF membrane by wet transfer.
  • 5. The PVDF membrane was incubated with 25 ml of blocking buffer [Tris Buffered Saline, pH 7.4 plus 0.1% TW20 (TBST)] containing 5% (W/V) BSA at room temperature for 1 hour.
  • 6. The membrane was rinsed with TBST once.
  • 7. The membrane was immersed with the protein side up in the primary antibody solution in TBST containing 5% (W/V) BSA and incubated for 24 hours at 4°C.
  • 8. The membrane was rinsed in TBST thrice for 5 minutes each.
  • 9. The membrane was incubated in the HRP-conjugated secondary antibody solution in TBST containing 5% (W/V) BSA and incubated for 1 hour at room temperature (~26°C) with gentle agitation.
  • 10. The membrane was rinsed thrice TBST thrice for 5 minutes each.
  • 11. The membrane was rinsed in TBS twice for 30 seconds each.
  • 12. Signals were detected with ECL-2 Substrate. The blot was scanned for 1 minute.
  • 13. The membrane was rinsed three times TBST.
  • 14. Incubated in Acidic Glycine Stripping Buffer at room temperature with gentle agitation for 3 times, 10 minutes each.
  • 15. The membrane was washed in TBST 2 times for 10 minutes each.
  • 16. Repeated Steps 5-12 with the loading control antibody (for Anti-actin) and its matching secondary antibody.
Anmerkungen None reported
Validierungsbilder
Western Blotting validation image for anti-Glutathione S Transferase (GST) antibody (ABIN1998462) Figure 1. Probing of cell extracts with GST Tag (upper panel) or loading control Acti...
Immunogen Recombinant GST protein
Klon 6C10G4
Isotyp IgG1
Spezifität Recognize GST Tag in fusion proteins.
No cross-reactivity with E.coli cell lysate in ELISA.
Keine Kreuzreaktivität Escherichia coli (E. coli)
Produktmerkmale This antibody was produced from a hybridoma resulting from the fusion of a mouse myeloma with B cells obtained from a mouse immunized with purified, recombinant Glutathione S-transferase (GST). The IgG fraction of the cell culture supernatant was purified by Protein A affinity chromatography.
Reinigung purified by Protein A affinity chromatography
Sterilität 0.2 μm filtered
Andere Bezeichnung GST (GST Antibody Abstract)
Hintergrund Genetic engineers have used glutathione S-transferase to create the GST gene fusion system. This system is used to purify and detect proteins of interest. In a GST gene fusion system, the GST sequence is incorporated into an expression vector alongside the gene sequence encoding the protein of interest. Induction of protein expression from the vector's promoter results in expression of a fusion protein: the protein of interest fused to the GST protein. This GST-fusion protein can then be purified from cells via its high affinity for glutathione. GST is commonly used to create fusion proteins. The tag has the size of 220 amino acids (roughly 26 KDa), which, compared to other tags like the myc- or the FLAG-tag, is quite big. However, many commercially-available sources of GST-tagged plasmids include a thrombin domain for cleavage of the GST tag during protein purification.
Forschungsgebiet Metabolism, Immunology
Applikationshinweise WB: 1/1000-1/10000
IP: 1-4 μL/mg of lysate
Beschränkungen Nur für Forschungszwecke einsetzbar
Format Liquid
Buffer 0.2 μm filtered solution in PBS
Konservierungsmittel Without preservative
Handhabung Avoid repeated freeze-thaw cycles.
Lagerung 4 °C,-20 °C,-80 °C
Informationen zur Lagerung This antibody can be stored at 2°C-8°C for one month without detectable loss of activity. Antibody products are stable for twelve months from date of receipt when stored at -20°C to -80°C. Preservative-Free.
Sodium azide is recommended to avoid contamination (final concentration 0.05%-0.1%). It is toxic to cells and should be disposed of properly. Avoid repeated freeze-thaw cycles.
Haltbarkeit 12 months
Bilder des Herstellers
Western Blotting (WB) image for anti-Glutathione S Transferase (GST) antibody (ABIN1998461) anti-Glutathione S Transferase (GST) antibody
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