GST-Tag 抗体

ABIN3045984 产品详细信息, 供应商: Log in to see
抗原
适用
Schistosoma japonicum
77
46
25
14
8
6
1
1
宿主
小鼠
118
28
16
3
1
1
克隆类型 (克隆位点)
单克隆 ()
标记
非结合性
11
6
6
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)
152
80
19
17
14
7
4
2
1
1
1
1
选项
Supplier
Log in to see
Supplier Product No.
Log in to see
'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
  • Supplier: Log in to see
  • Supplier catalog number: Log in to see
  • Lot number: 1214
  • Dilution: 1:1000
Secondary Antibody
  • Antibody: HRP labeled secondary anti-mouse antibody
  • Supplier: Log in to see
  • 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
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
抗原 GST-Tag
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
您还需要查找其他产品吗?