GST-Tag antibody

Details for Product No. ABIN3045984, Supplier: Log in to see
Antigen
Reactivity
Schistosoma japonicum
78
55
26
14
8
3
1
1
1
Host
Mouse
121
23
14
4
1
Clonality (Clone)
Monoclonal ()
Conjugate
Un-conjugated
11
7
7
6
6
6
6
5
5
5
5
5
5
5
5
5
5
5
2
1
1
1
1
1
Application
Chromatin Immunoprecipitation (ChIP), Immunoprecipitation (IP), Immunohistochemistry (IHC), Western Blotting (WB)
150
80
23
19
17
10
6
2
1
1
1
Options
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'Independent Validation' Badge
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.
Primary Antibody
  • Antibody: Mouse anti-GST monoclonal antibody
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  • Supplier catalog number: Log in to see
  • Lot number: 1214
  • Dilution: 1:1000
Secondary Antibody
  • Antibody: HRP labeled secondary anti-mouse antibody
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  • Supplier catalog number: Log in to see
  • Lot number: 390630
  • Dilution: 1:2000
Controls
  • Positive control: BL21 bacteria cells were transformed with an in-house GST expression vector pGST
  • Negative control: Empty vector cell lysate
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).
Validation Images
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
Clone 5C12
Isotype IgG1
Characteristics The GST tag issued in many protocols
Purification purified form hybridoma supernatant
Alternative Name GST Tag
Molecular Weight 26 kDa
UniProt P08515
Application Notes WB 0.1-1.0ug/ml
Restrictions For Research Use only
Format Liquid
Concentration 1 mg/mL
Buffer PBS pH7.2
Storage -20 °C