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GST-Tag 抗体

ABIN3045984 产品详细信息, 供应商: Log in to see
抗原
适用
Schistosoma japonicum
78
55
26
14
8
3
1
1
1
宿主
小鼠
121
23
14
4
1
克隆类型 (克隆位点)
单克隆 ()
标记
非结合性
11
7
7
6
6
6
6
5
5
5
5
5
5
5
5
5
5
5
2
1
1
1
1
1
应用范围
Chromatin Immunoprecipitation (ChIP), Immunoprecipitation (IP), Immunohistochemistry (IHC), Western Blotting (WB)
150
80
23
19
17
10
6
2
1
1
1
Supplier
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Supplier Product No.
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Available images

'独立验证'标志
抗原 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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  • Lot number: 1214
  • Dilution: 1:1000
Secondary Antibody
  • Antibody: HRP labeled secondary anti-mouse antibody
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  • 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
实验流程
  • **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...
免疫原 GST
克隆位点 5C12
亚型 IgG1
产品特性 The GST tag issued in many protocols
纯化方法 purified form hybridoma supernatant
别名 GST Tag
分子量 26 kDa
UniProt P08515
应用备注 WB 0.1-1.0ug/ml
限制 仅限研究用
状态 Liquid
浓度 1 mg/mL
缓冲液 PBS pH7.2
储存条件 -20 °C