货号: | EZIP04-5 |
供应商: | 上海万生昊天生物技术有限公司 |
规格: | 5ml |
The V5 epitope tag is derived from a small epitope (Pk) present on the P and V proteins of the paramyxovirus of simian virus 5 (SV5). The V5 tag antibody can be helpful in detecting the recombinant proteins, some of which include transmembrane and secreted proteins fusion protein. This V5-tag antibody is prepared by immunizing synthetic peptide GKPIPNPLLGLDST coupled to KLH.
Isotype Mouse IgG1
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 V5 fusion proteins with Anti-V5 tag antibody Affinity Gel.
1. Thoroughly suspend the Anti-V5 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-V5 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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