GST antibody (Glutathione S Transferase)

Details for Product anti-GST Antibody No. ABIN1998461, Supplier: Log in to see
Antigen
  • GST
  • GST 13-13
  • GST13
  • GST13-13
  • GSTK1-1
  • hGSTK1
  • GSTS
  • GSTS1-1
  • PGD2
  • PGDS
  • H-PGDS
  • Ptgds2
  • 1500002K10Rik
  • Gst
  • glutathione S-transferase kappa 1
  • hematopoietic prostaglandin D synthase
  • microsomal glutathione S-transferase 1
  • GSTK1
  • HPGDS
  • Hpgds
  • Mgst1
Reactivity
Schistosoma japonicum
291
85
35
26
11
9
4
3
2
2
1
1
1
Host
Mouse
201
149
92
5
5
4
2
1
Clonality (Clone)
Monoclonal ()
Conjugate
This GST antibody is un-conjugated
32
29
25
13
7
6
5
4
4
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
Application
Immunoprecipitation (IP), Western Blotting (WB)
357
216
95
86
41
31
17
13
12
11
9
9
9
9
7
6
5
3
3
2
2
2
1
1
1
1
1
1
1
1
Options
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'Independent Validation' Badge
Antigen Glutathione S Transferase (GST)
Lot Number HD04FE1006
Method validated Western Blotting
Positive Control GST- AcNFkBp(C-terminal) 293-Lysate
Negative Control Empty vector- 293 Lysate
Notes A strong specific band was observed in the positive control at the expected size (~82 kDa) that is not observed in the negative control.
Primary Antibody
  • Antigen: Glutathione S Transferase (GST) protein
  • Catalog number: ABIN1998462
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  • Supplier catalog number: Log in to see
  • Lot number: HD04FE1006
  • Antibody Dilution: 1:7000
Secondary Antibody
  • Antigen: Goat Anti-Mouse IgG (H + L)-HRP Conjugate
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  • Catalog number: Log in to see
  • Lot number: N/A
  • Antibody Dilution: 1:15,000
Controls
  • Positive control: GST- AcNFkBp(C-terminal) 293-Lysate
  • Negative control: Empty vector- 293 Lysate
Protocol
  • 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.
Experimental Notes None reported
Validation Images
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
Clone 6C10G4
Isotype IgG1
Specificity Recognize GST Tag in fusion proteins.
No cross-reactivity with E.coli cell lysate in ELISA.
No Cross-Reactivity Escherichia coli (E. coli)
Characteristics 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.
Purification purified by Protein A affinity chromatography
Sterility 0.2 μm filtered
Alternative Name GST (GST Antibody Abstract)
Background 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.
Research Area Metabolism, Immunology
Application Notes WB: 1/1000-1/10000
IP: 1-4 μL/mg of lysate
Restrictions For Research Use only
Format Liquid
Buffer 0.2 μm filtered solution in PBS
Preservative Without preservative
Handling Advice Avoid repeated freeze-thaw cycles.
Storage 4 °C,-20 °C,-80 °C
Storage Comment 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.
Expiry Date 12 months
Supplier Images
Western Blotting (WB) image for anti-GST antibody (Glutathione S Transferase) (ABIN1998461) anti-Glutathione S Transferase (GST) antibody