货号: | EZIP06-1 |
供应商: | 上海万生昊天生物技术有限公司 |
规格: | 1ml |
A GST-tag is often used to separate and purify proteins that contain the GST-fusion protein. The tag is 220 amino acids (roughly 26 KDa), in size is quite large. It is fused to the N-terminus of a protein. However, many commercially available sources of GST-tagged plasmids include a thrombin domain for cleavage of the GST tag during protein purification.
Isotype Mouse IgG2b
Form Slurry
Source Purified antibody was conjugated to gel on optimal condition.
Concentration 50% (v/v) in PBS pH7.4, 0.02% Sodium Azide, 55% glycerol.
Storage -20℃
Applications IP, protein purification
Specificity All species
Procedure
Isolation of GST fusion proteins with Anti-GST tag antibody Affinity Gel.
1. Thoroughly suspend the Anti-GST Affinity Gel in the vial, in order to make a uniform suspension of the resin. Quickly transfer 10μl of the gel suspension (about 5μl of packed gel volume) to a fresh tube.
2. Add 0.6ml TBS. Thoroughly suspend the Anti-GST Affinity Gel. Centrifuge the resin at 10000 rpm for 30 seconds. Remove the supernatant carefully. Be sure that most of the wash buffer is removed and no resin is discarded. This step should be repeated for 3-4 times.
3. Add 500μl of E .coli periplasmic extracts to the washed resin.
4. Agitate all samples gently for 2 hours at 4℃.
5. Centrifuge the resin for 30 seconds at 10000 rpm. Remove the supernatants to a fresh tube.
6. Wash the resin with 0.5ml TBS until the OD280 of the supernatant liquid<0.05.
7. Add 10μl of 2×sample buffer to each sample (the resin and the supernatant). Boil the samples for 5 minutes.
8. Load 4μl of each sample on SDS-PAGE.
FOR RESEARCH USE ONLY.
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