GST-Tag 抗体
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北京 101111
Quick Overview for GST-Tag 抗体 (ABIN5540371)
抗原
适用
宿主
克隆类型
标记
应用范围
克隆位点
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特异性
- This antibody recognizes recombinant GST-Tag specifically.
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纯化方法
- Protein A sepharose
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免疫原
- Recombinant GST protein
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亚型
- IgG2b
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anti-GST-Tag antibody
WB 宿主: 小鼠 Monoclonal 9AT106 unconjugated
anti-GST-Tag antibodyWB 宿主: 兔 Polyclonal RB2165 unconjugated
anti-GST-Tag antibodyWB, 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 宿主: 小鼠 Monoclonal 25GB7025 unconjugated
anti-GST-Tag (AA 1-201) antibodyWB, IP 宿主: 兔 Monoclonal BLRE05M unconjugated Recombinant Antibody
anti-GST-Tag antibodyWB 宿主: 小鼠 Monoclonal 25GB7045 unconjugated
anti-GST-Tag antibodyWB, IP 宿主: 小鼠 Monoclonal S-tag-05 unconjugated
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应用备注
- Western blot: 1 μg/mL. Immunoprecipitation: 5 μg/100 μL of E. coli extracts containing GST - Tag fusion protein.
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实验流程
- SDS PAGE & Western Blotting 1) Boil the sample for 3~5 minutes and centrifuge at 12,000 x g for a minute. Other methods may be suitable. 2) Resolve 10 μ l of sample by SDS - polyacrylamide gel electrophoresis (See the manufacture's manual for electrophoresis c onditions). 3) Transfer to a polyvinylidene difluoride (PVDF) membrane at 1 mA/cm 2 for 1 hour in a semi - dry transfer system. (Transfer Buffer: 25 mM Tris, 190 mM glycine, 20 % MeOH). See the manufacture's manual for specific transfer procedure. 4) The transferred proteins can be visualized by staining the membrane for 1 minute with Ponceau. Rinse the membrane with PBS. 5) Non - specific binding sites are blocked by immersing the membrane in 5 % Skim Milk/PBS/0.05 % Tween 20 for 1 hr.at room temperat ure (20 - 25 o C) or for overnight at 4 o C. 6) Incubate in primary antibody diluted as suggested in the APPLICATIONS for 1 hour at room temperature(20 - 25 o C). (The optimal antibody concentration will depend on the experimental conditions.) 7) Wash the membrane 3 times with PBS containing 0.05 % Tween20 for 5~10 minutes each. 8) Incubate the membrane with secondary antibody (1:10,000 diluted horseradish peroxidase conjugated anti mouse IgG (H+L) antibody in PBS containing 0.05 % Tween 20 for 45 minutes at ro om temperature (20 - 25 o C). 9) Wash the membrane 3 times with PBS containing 0.05 % Tween20 for 5~10 minutes each. 10) Wipe excess buffer from the membrane, then incubate it with appropriate chemiluminescence reagents for 1 minute. Remove extra reagent from the membra ne by dabbing with a paper towel, and seal it in plastic wrap. 11) Expose to X - ray film in a dark room for 30 seconds. Develop the film as usual. The conditions for exposure and development may vary. Immunoprecipitation 1) Add 5 μ g of the antibody to 100 μ l of E. Coli extracts containing GST - Tag fusion protein. Mix well and incubate with gentle agitaiton for 30 - 120 minutes at 4 o C. 2) Add 20 μ l of 50 % Protein A - agarose beads. Mix well and incubate with gentle agitation for 1 hour at 4 o C. 3) Wash the beads 4 times with ice - cold Lysis buffer (50 mM HEPES, pH 7.3, 250 mM NaCl, 0.2 % N P - 40, 5 mM EDTA, 10 % glycerol), then centrifuge the tube at 2,500 x g for 10 seconds to remove the buffer. 4) Resuspend the beads in 40 μ l of Laemmli SDS PAGE sampl e buffer, boil for 3 - 5 minutes, and centrifuge for 5 minutes. Apply 20 μ l/lane for the SDS - PAGE analysis (See SDS - PAGE & Western blotting.
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限制
- 仅限研究用
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状态
- Liquid
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缓冲液
- PBS containing 50 % glycerol. No preservative is contained.
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储存液
- Azide free
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储存条件
- -20 °C
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储存方法
- Upon receipt, store undiluted (in aliquots) at -20°C. Avoid repeated freezing and thawing. Shelf life: One year from despatch.
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- GST-Tag
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物质类
- Tag
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背景
- Expression vectors containing a protein and a tag protein are commonly used. GST - Tag fusion protein expression sys tem is preferably used in various laboratories, because its simple protein purification step by an affinity chromatograpy. This specific antibody for GST - Tag fusion protein is useful tools for monitoring of the fusion protein expression and affinity purifi cation.
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UniProt
- P08515
抗原
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