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Flu-P1 (Lyophilized) |
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Description:
Flu-P1 is a synthetic peptide containing an N-terminal cysteine and a fluorescein-labeled lysine. This peptide can be used in conjunction with the IMPACT Kit (NEB #E6901) to site specifically incorporate a fluorescein moiety into a bacterially-expressed protein. The Flu-P1 peptide ligates to the C-terminus of proteins containing a C-terminal thioester (1,2,3). This procedure is termed intein-mediated protein ligation (IPL). The thioester-tagged proteins are purified using the IMPACT Kit in which, preferably, 2-mercaptoethanesulfonic acid (MESNA) was used to induce intein cleavage. This peptide can also be used in conjunction with the Peptide Carrier Kit (NEB# E6600) to label Carrier Protein 39 and 27 for western blot analysis.
The peptide sequence is NH2-CDPEK(Fl)DS-COOH. The K(Fl) represents a lysine with a fluorescein covalently attached to its ε-amino group.
Source:
An HPLC purified synthetic peptide.
Applications:- Site specifically labeling the C-terminus of a protein with fluorescein.
Reagents Supplied:
IPL Reaction Buffer (5X)
Properties & Usage
Molecular Weight:
Theoretical: 1,182 daltons
Reaction & Storage Conditions
Reaction Conditions:
1X IPL Reaction Buffer
Incubate at
25°C.
1X IPL Reaction Buffer:
100 mM Tris-HCl
100 mM NaCl
50 mM MESNA
pH 8.5 @ 25°C
Storage Temperature:
-20°C
Notes
Usage notes:- Upon first use resuspend the peptide in 50 µl of 5X IPL Reaction Buffer. Separate into 10 µl aliquots and store at -70°C for future use.
Figure 1: Intein-mediated Protein Ligation
References
- Evans, T. C. et al. (1998) Protein Sci., 2254-2264.
- Muir, T. W. (1998) Proc. Natl. Acad. Sci. USA, 95, 6705-6710.
- Xu, M.-Q and Evans, T. C., Jr. (2001) Methods, 24, 257-277.
Companion Products
Bio-P1 (Lyophilized)
Peptide-Carrier™ Kit
TransPass™ P Protein Transfection Reagent
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