GST-Tag 抗体
(1 validation)Quick Overview for GST-Tag 抗体 (ABIN3045984)
抗原
适用
宿主
克隆类型
标记
应用范围
克隆位点
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产品特性
- The GST tag issued in many protocols
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纯化方法
- purified form hybridoma supernatant
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免疫原
- GST
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亚型
- IgG1
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anti-GST-Tag antibody
适用: Schistosoma japonicum WB, ELISA 宿主: 小鼠 Monoclonal 9AT106 unconjugated
anti-GST-Tag (AA 1-218) antibody适用: Various Species WB, IP, DB 宿主: 小鼠 Monoclonal C9 unconjugated
anti-GST-Tag (AA 1-218) antibody适用: Various Species WB, IP, DB 宿主: 兔 Polyclonal unconjugated
anti-GST-Tag antibodyWB, IP 宿主: 小鼠 Monoclonal S-tag-05 unconjugated
anti-GST-Tag antibodyWB 宿主: 小鼠 Monoclonal 9AT106 unconjugated
anti-GST-Tag antibodyWB 宿主: 兔 Polyclonal RB2165 unconjugated
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应用备注
- WB 0.1-1.0ug/ml
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限制
- 仅限研究用
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- by
- Celplor LLC
- No.
- #029860
- 日期
- 2016.05.19
- 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
- Lot number: 1214
- Dilution: 1:1000
- Secondary Antibody
- Antibody: HRP labeled secondary anti-mouse antibody
- Lot number: 390630
- Dilution: 1:2000
- Full 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).
- Experimental Notes
生效 #029860 (Western Blotting)!['独立验证'标志]( "成功验证")
!['独立验证'标志]( "成功验证")
Validation Images![Figure 1: GST content of BL21-pGST(+IPTG) is 12% of total protein. GST content in IPTG induced samples was analyzed by Quantity One software (Bio-Rad).]()
Figure 1: GST content of BL21-pGST(+IPTG) is 12% of total protein. GST content in IPTG induced samples was analyzed by Quantity One software (Bio-Rad).
![Figure 2: Western blot for mouse anti-GST monoclonal antibody. The first lane shows the empty vector cell lysate (negative control). This is followed by a 0.01, 0.1, and 1 ug of total protein from BL21-pGST, with (Lanes 2, 3, and 4) or without (Lanes 5, 6, and 7) IPTG induction, respectively.
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. The higher than expected molecular weight may be the result GST dimers or aggregates (Riley et al. Protein Engineering vol.9 no.2 pp.223-23O, 1996).]()
Figure 2: Western blot for mouse anti-GST monoclonal antibody. The first lane shows the empty vector cell lysate (negative control). This is followed by a 0.01, 0.1, and 1 ug of total protein from BL21-pGST, with (Lanes 2, 3, and 4) or without (Lanes 5, 6, and 7) IPTG induction, respectively.
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. The higher than expected molecular weight may be the result GST dimers or aggregates (Riley et al. Protein Engineering vol.9 no.2 pp.223-23O, 1996).
Full Methods -
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状态
- Liquid
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浓度
- 1 mg/mL
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缓冲液
- PBS pH7.2
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储存条件
- -20 °C
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- GST-Tag
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别名
- GST Tag
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物质类
- Tag
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分子量
- 26 kDa
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UniProt
- P08515
抗原
-
