Thermo Scientific Pierce Crosslinking Technical Handbook

Crosslinking Applications
Label transfer reagents
Thermo Scientific Pierce Bifunctional label transfer reagents
Heterobifunctional, photoreactive, thiol-cleavable label transfer
reagents enable the tagging of a prey protein. The photolysis wave
lengths for these reagents are in the range between 320-400 nm,
limiting damage to biomolecules by irradiation.
Ordering Information
Product # Description Structure Features Ref. Pkg. Size
27720 APDP
N-[4-(p-Azido-salicylamido)
butyl]-3´-(2´-pyridyldithio)
propionamide
M.W. 446.55
Spacer Arm 21.0 Å
N
S
S
O
H
N
N
H
O
HO
N
N +
N –
• Radioiodinatable between –N 3
and –OH group of phenyl ring
• –SH-reactive
• Reaction monitored at 343 nm
• Membrane permeable
1-5 50 mg
33030 Sulfo-SAED
Sulfosuccinimidyl 2-(7-
azido-4-methylcoumarin-
3-acetamido)ethyl-1,3
dithiopropionate
M.W. 621.6
Spacer Arm 23.6 Å
27719 Sulfo-SFAD
Sulfosuccinimidyl(perfluoroazidobenzamido)ethyl-1,3´dithiopropionate
M.W. 597.48
Spacer Arm 14.6 Å
O
N
O
O
Na +– O
S O
O
Na + O –
S
O
O
O
O
N
O
O
O
S
S
H
N
SAED
M.W. 621.60
Spacer Arm 23.6 Å
S
S
SFAD
M.W. 597.48
Spacer Arm 14.6 Å
O
O O
N
N +
N
H
O F
F
F
F
N –
N
N +
N –
• Water-soluble
• Amine-reactive
• Photoreactive
• Prey protein tracked by
fluorescence
• Ex: 345-350 nm, Em: 440-460 nm
• No radiolabeling required
• AMCA moiety exhibits large
Stokes shift
• Improved photoconjugation
efficiency
• Photolyzes at 320 nm
• Label transfer monitored by
19
F NMR
• Water-soluble
• Cleavable
• Amine-reactive
16,17 5 mg
9,10,
14,15
50 mg
Compatible Pierce products for addition of 125 I to APDP.
Ordering Information
Product # Description Pkg. Size
28601 Pierce Pre-Coated Iodination Tubes
(12 mm x 75 mm glass test tubes coated with
50 µg Pierce Iodination Reagent in 100 µl volume)
28665 Pierce Iodination Beads
(N-Chloro-benzenesulfonamide modified
non-porous polystyrene beads)
28666 Pierce Iodination Beads
(N-Chloro-benzenesulfonamide modified
non-porous polystyrene beads)
28600 Pierce Iodination Reagent
(1,3,4,6-Tetrachloro-3α,6α-diphenylglycoluril)
10 tubes/
pkg.
50/pkg.
250/pkg.
1 g
References
1. Dey, D., et al. (1998). J. Biol. Chem. 273(3), 1670-1676.
2. Sato, K., et al. (1997). J. Biol. Chem. 272(9), 5880-5886.
3. Gao, A.G. and Frazier, W.A. (1994). J.Biol. Chem. 269(47), 29650-29657.
4. Traut, R.R., et al. (1989). Protein Function, A Practical Approach,
Oxford:IRL Press, p. 101.
5. Lam, Y.A., et al. (2002). Nature 416 (April 19), 763-766.
6. Felin, M., et al. (1997). Glycobiology 7(1), 23-29.
7. Lala, A., et al. (1996). Pept. Res. 9(2), 58-60.
8. Zhang, Q. and Mosher, D.F. (1996). J. Biol. Chem. 271(52), 33284-33292.
9. Chattopadhyay, A., et al. (1992). J. Biol. Chem. 267, 12323-12329.
10. Smith, J.W., et al. (1990). J. Biol. Chem. 265(21), 12267-12271.
11. Smith, J.W. and Cheresh, D.A. (1990). J. Biol. Chem. 265(4), 2168-2172.
12. Ghinea, N., et al. (1989). J. Biol. Chem. 264(9), 4755-4758.
13. Shephard, E.G., et al. (1988). Anal. Biochem. 186, 306-313.
14. Pandurangi, R.S., et al. (1998). Bioorg. Chem. 26(4), 201-212.
15. Pandurangi, R.S., et al. (1997). Photochem. Photobiol. 65(2), 208-221.
16. Thevenin, B., et al. (1991). Biophys. J. 59, 358a.
17. Kang, J., et al. (1991). Biophys. J. 59, 249a.
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For more information, or to download product instructions, visit www.thermo.com/pierce