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CuAAC Reaction Ligand Test Kit (THPTA & BTTAA based)

Cat. No. Amount Price (EUR) Buy / Note
CLK-075 1 kit 119,65 Add to Basket/Quote Add to Notepad

For research use only!

Shipping: shipped at ambient temperature

Storage Conditions: store at 4 °C
Short term exposure (up to 1 week cumulative) to ambient temperature possible.

Shelf Life: 12 months after date of delivery

Description:
The CuAAC Reaction Ligand Test Kit is suitable to perform Copper (Cu(I))-catalyzed Azide-Alkyne Click chemistry reactions (CuAAC) with Azide- or Alkyne- modified biomolecules and cells containing metabolically functionalized Alkyne- or Azide-modified biomolecules. It allows a direct comparison of BTTAA and THPTA to find the most suitable Cu(I)-stabilizing ligand for your experimental set-up.
1 Kit provides sufficient amounts to perform 25 CuAAC experiments each with THPTA and BTTAA à 200 μl using 2 mM CuSO4 (copper source), 10 mM THPTA or 10 mM BTTAA (Cu(I)-stabilizing ligands) and 100 mM Na-Ascorbate (reduction reagent) in 100 mM Na-Phosphate reaction buffer.

Content:
Copper source:
1 x 10 mg CuSO4 (M = 159.6 g/mol), #CLK-MI004)

Cu(I) stabilizing ligands:
1 x 25 mg THPTA (M= 434.5 g/mol, #CLK-1010)
1 x 25 mg BTTAA (M= 430.5 g/mol, #CLK-067)

Reduction Reagent:
2 x 200 mg Na-Ascorbate (M = 119.3 g/mol, #CLK-MI005)

Reaction Buffer:
1 x 30 ml sterile 100 mM Na-Phosphate Buffer, pH 7

10 ml sterile ddH2O

Materials required but not provided:

Alkyne-or Azide-functionalized substrates e.g.
a) fixed and permeabilized cells containing metabolically functionalized Alkyne- or Azide-modified biomolecules.
b) cell lysate containing metabolically functionalized Alkyne- or Azide-modified proteins.
c) Alkyne- or Azide-functionalized biomolecules such as DNA, RNA or oligonucleotides.

(Picolyl)-Azide or Alkyne detection reagent and appropriate solvent (e.g. DMSO)

For labeling of fixed and permeabilized cells:
Washing solutions e.g. PBS containing 3% BSA
Fixation solution e.g. PBS containing 3.7% formaldehyd
Permeabilization solution e.g. PBS containing 0.5% Triton X-100
Mounting medium for imaging
Additional labeling reagent such as nuclear stain or antibody

Selected References:
[1] Presolski et al. (2011) Copper-Catalyzed Azide-Alkyne Click Chemistry for Bioconjugation. Current Protocols in Chemical Biology 3:153.
[2] Hong et al. (2011) Analysis and Optimization of Copper-Catalyzed Azide-Alkyne Cycloaddition for Bioconjugation. Angew. Chem. Int. Ed. 48:9879.
[3] Besanceney-Webler et al. (2011) Increasing the Efficiacy of Bioorthogonal Click Reactions for Bioconjugation: A Comparative Study. Angew. Chem. Int. Ed. 50:8051.
[4] Uttamapinant et al. (2012) Fast, Cell-Compatible Click Chemistry with Copper-Chelating Azides for Biomolecular Labeling. Angew. Chem. Int. Ed. 51:5852.