TurboGFP has been a widely used fluorescent marker and in many cases adding a TurboGFP tag will not affect function or localization of the protein. However, in some cases the tag could interfere with function or localization of the protein by affecting protein folding, for example. To help prevent this from occurring, we include a short linker peptide between the target protein and the TurboGFP tag.
We insert the desired tag using a homology-independent strategy we published in Nature Communications. In this strategy the target gene is cleaved at the site where the tag will be integrated using Cas9 programmed with a gRNA. The tag is the inserted via non-homologous end joining at the site where the gRNA cleaved. For more information, please see: A generic strategy for CRISPR-Cas9-mediated gene tagging doi: 10.1038/ncomms10237
Since the TurboGFP tag is incorporated at the endogenous locus, the intensity of the signal depends on the expression level of the gene being tagged.
One strategy to amplify the signal is to fix the cells and do an immune-fluorescence assay using a TurboGFP antibody.
Yes. A vial of Hap1 WT cells will be included at no additional cost.
TurboGFP has high photostability and should not be easily bleached.
No, the tagging plasmid is not provided as a product.
Yes, Turbo RFP, NanoLuc and Halo tags are also available.
The editing process ensures that all proteins are tagged, there are no untagged alleles.
Yes, simply complete our request form for the GFP visualization protocol
We recommend to keep the cells in culture for approximately 15 passages.
The intensity of the tag itself should not change in culture over time. However, if expression levels of the tagged gene change, the intensity of TurboGFP will change accordingly.
TurboGFP has a rapid generation of fluorescent signal in living cells and should be visible as soon after the target gene is expressed.
Live cell imaging, studying the function and localization of a protein, western blotting, protein pull down, affinity chromatography, immunocytochemistry and flow cytometry.
Molecular weight, kDa 26
Polypeptide length, aa 232
The sequence does not include a start or stop codon.
|Excitation maximum, nm||482|
|Emission maximum, nm||502|
The cells can be cultured the same as wildtype Hap1 cells.
Individual antibodies can require specific protocols, so it is advisable to follow the protocol from the antibody manufacturer.
TurboGFP can be used in multicolor labeling applications with blue, true-yellow, red, and far-red fluorescent dyes.