货号: | EZIP01-10 |
供应商: | 上海万生昊天生物技术有限公司 |
规格: | 10ml |
FLAG-tag, or FLAG octapeptide, or FLAG epitope, is a polypeptide protein tag that can be added to a protein using recombinant DNA technology, having the sequence motif DYKDDDDK. It represents an early example of a nanotechnology device, in that it can be attached to a protein like a molecular handle, then used to manipulate the protein, and subsequently detached. It has been used for studying proteins in living cells and for protein purification by affinity chromatography.
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 Flag fusion proteins with Anti-Flag tag antibody Affinity Gel.
1. Thoroughly suspend the Anti-Flag 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-Flag 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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温馨提示:不可用于临床治疗。