货号: | EZIP05-10 |
供应商: | 上海万生昊天生物技术有限公司 |
规格: | 10ml |
A polyhistidine-tag is an amino acid motif in proteins that consists of at least six histidine (His) residues, often at the N- or C-terminus of the protein. It is also known as hexa histidine-tag, 6xHis-tag, His6 tag and by the trademarked name His-tag. Polyhistidine-tags are often used for affinity purification of genetically modified proteins.
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 His fusion proteins with Anti-His tag antibody Affinity Gel.
1. Thoroughly suspend the Anti-His 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-His 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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