PTAD-Azide

C₁₀H₁₀N₆O₃

262.22

MFCD25976544

329772652

NA.22

PTAD-Azide (4-(4-(2-Azidoethoxy)phenyl)-1,2,4-triazolidine-3,5-dione) is a 1,2,4-triazolidine-3,5-dione derivative. It can be prepared from ethyl hydrazinecarboxylate.

PTAD-Azide is a selective crosslinking reagent that has one end for reacting with tyrosine and the other end for presenting an azide. After bioconjugation to tyrosine, the azide can be reacted with an alkyne through the Cu(I)-catalyzed click chemistry reaction or with a cyclooctyne in a copper-free reaction. This reagent has been shown to selectively introduce poly(ethylene glycol) or PEG chains onto proteins with surface exposed tyrosine residues. PTAD-Azide has also been used in the formation of antibody-drug conjugates. This reagent is compatible with different buffer systems such as PBS, Tris and mixed PBS/Tris buffer (preferred). The linkage with tyrosine has been shown to be stable to pH and temperature extremes as well as blood plasma.

Note: PTAD-Azide must be first activated by stirring in a 1:0.98 molar ratio with 1,3-dibromo-5,5-dimethylhydantoin (product # 157902). Activation is evident upon solution color change from colorless to deep red and the activated reagent should be used immediately.


General Procedure for Protein Modification with PTAD.

Part 1: PTAD activation

• Mix together 1:0.98 molar equivalents of unactivated PTAD to 1,3-dibromo-5,5-dimethylhydantoin (product # 157902) in organic solvent (preferred solvents are DMF or acetonitrile, avoid using DMSO)
• Color change is observed from colorless/pale yellow to deep red (approximately 5 min of mixing).
• After the solution turns red, store the now activated reagent on ice and use for protein modification within 30 min.

Part 2: Protein modification

• Add protein solution in mixed phosphate/Tris buffer or Tris buffer (pH should be 6 - 9) to the eppendorf tube (or other vial) containing the activated PTAD reagent prepared above and mix gently at room temperature for up to 30 min. Preferably use 10-fold molar excess of reagent relative to protein. Use protein at a minimum concentration of 1 mg/ml (higher concentrations are preferred for enhanced labeling).
• Remove excess unreacted PTAD by gel filtration.

PTAD-Azide may be used in the preparation of 4-(4-(2-azidoethoxy)phenyl)-3H-1,2,4-triazole-3,5(4H)-dione and aplaviroc-urazole. This urazole was recently demonstrated in the traceless, chemoselective labeling of peptides and proteins through electrochemical tyrosine-click (e-Y-CLICK) chemistry.

Electrochemically Promoted Tyrosine-Click-Chemistry for Protein Labeling

95%

powder or crystals

reagent type: cross-linking reagent

2-8℃

O=C(NNC1=O)N1C2=CC=C(OCCN=[N+]=[N-])C=C2

1S/C10H10N6O3/c11-15-12-5-6-19-8-3-1-7(2-4-8)16-9(17)13-14-10(16)18/h1-4H,5-6H2,(H,13,17)(H,14,18)

MHGMHPVYCVQIET-UHFFFAOYSA-N

50 mg in glass bottle

13 - Non Combustible Solids

WGK 1