What sequence is a His tag?
The features
| Features of the His-tag: | |
|---|---|
| Amino Acid Sequence | HHHHHH (4-10) |
| DNA Sequence | 5′-CAT CAC CAT CAC CAT CAC-3′ |
| Size | 840.8 Da (His6) |
| Compatibility to recombinant proteins | Can be added either to the N- or C-Terminus of a protein |
How big is His tag?
∼2.5 kDa
His-tags, due to their relatively small size (∼2.5 kDa), are not believed to significantly interfere with the function and structure of a majority of proteins.
How do you detect his-tagged proteins?
Check total protein content of the gel by staining the gel with a total protein stain. Load at least 1 picomole of the His-tagged fusion protein for detection. Make sure the His-tag is in frame and the protein is expressed properly. Be sure to wash the gel twice with 20 mM phosphate buffer.
Does his tag affect protein activity?
Yes it looks that his-Tag can affect the protein activity as I expressed my 10KD protein in pET32a, and then check my protein activity in fact no valuable effect was observed by the expressed protein.
What is the size of His tag?
His-tags. Molecular Weight: 0.2–1.6 kDa. 6x-His tag is 0.8 kDa. Tag location: C- or N- terminals, or internal.
Is His tag immunogenic?
Although the His-tag is a short and poorly immunogenic sequence, its attachment to a recombinant protein might mask immunologically important epitopes and reduce the potential immune response towards the target antigen, a potential impact that requires experimental evaluation [22].
How many kDa is a His-tag?
2.5 kDa
How heavy is His-tag?
His-tags. Molecular Weight: 0.2–1.6 kDa.
How do you identify a his-tag protein?
Detection of His-Tag proteins Epitope tagging is a common method employed for the identification and detection of proteins. In a similar manner, the His-Tag also acts as a specific site for a binding partner, such as an antibody with high specificity, so allowing easy detection of the recombinant protein of interest.
How many histidine residues are in a suitable tag sequence?
Suitable tag sequences are available free for commercial use; for example, MK (HQ)6 may be used for enhanced expression in E. coli and tag removal. The total number of histidine residues may vary in the tag from as low as two, to as high as 10 or more His residues.
How do you add a his-tag to a gene?
(A) The His-tag is added by inserting the DNA encoding a protein of interest in a vector that has the tag ready to fuse at the C-terminus. (B) The His-tag is added using primers containing the tag, after a PCR reaction the tag gets fused to the N-terminus of the gene.
What is a DNA tag?
His or GST). A tag is a short sequence of DNA that codes for a specific amino acid, which is frequently inserted into a target gene at the point of coding for expression at either the N or C terminal of the protein required.