Conjugation of nanoCLAMP(Cys) to a maleimide label


Preparation of labeled nanoCLAMP(Cys) by coupling a maleimide label to the nanoCLAMP's unique C-terminal cysteine. 


1 Collect 10 nmol lyophilized nanoCLAMP at the tube bottom with a quick spin
2 Add 100 ul NRB1 and 2 ul TCEP, 50 mM. Pipette gently until the pellet resuspends
3 Incubate the resuspended nanoCLAMP on ice for 30 minutes
4 Prepare a fresh solution of the maleimide label in "NRB1," DMF, DMSO or other suitable buffer
5 Add 10 ul of the maleimide label solution to the resuspended nanoCLAMP and mix by pipetting
6 Incubate the conjugation reaction in the dark overnight at Room Temperature
7 Remove unbound dye from the labeled nanoCLAMP by exchanging the buffer to "TBS pH 7.5" or equivalent with a Zeba™ Spin Desalting Column
8 Quantify dye incorporation in the desalted solution by measuring absorbance at the appropriate wavelength

Solution(s) Used
Use / Nickname Solution Usage Rate
For resuspending the nanoCLAMP and label NRB1 200 ul per reaction
For buffer exchange of the labeled nanoCLAMP TBS pH 7.5 1.2 ml per reaction

Outside Material(s) Used
Vendor Outside Material Catalog Number Usage Rate
ThermoFisher Scientific Zeba™ Spin Desalting Columns, 7K MWCO, 0.5 mL 89882 1 column per reaction