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 HOME > Protein Research > Protein assay |
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Maleimide Quantitation Kit |
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코 드 |
: 5523 |
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단 위 |
: 200 assay |
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공 급 원 |
: ABD Bioquest |
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가 격 |
: 문의 |
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Maleimide assay Kit
Amplite™ Fluorimetric Maleimide Quantitation Kit *Green Fluorescence*
A variety of crosslinking reagents with a maleimide group are widely used for crosslinking proteins to proteins or proteins to other biomolecules. There are few reagents or assay kits available for quantitate the number of maleimide groups that are introduced into the first protein. All the commercial kits have tedious protocols. Our kit uses a proprietary dye that has enhanced fluorescence upon reacting with a maleimide. The kit provides a sensitive, one-step fluorimetric method to detect as little as 10 picomole of maleimide in a 100 µL assay volume (100 nM in concentration). The assay is rapid and robust. It can be performed in a convenient 96-well or 384-well microtiter-plate format and easily adapted to automation with Ex/Em = 490 nm/520 nm.
Key Features
Broad Application : Can be used for quantifying maleimide group in a variety of molecules such as proteins.
Sensitive : Detect as low as 10 picomole of maleimide.
Continuous : Easily adapted to automation with no separation required.
Convenient : Formulated to have minimal hands-on time. No wash is required.
Non-Radioactive : No special requirements for waste treatment.
Kit Components
Component A : Maleimide Green™ 1 vial
Component B : Reaction Buffer 1 vial(500 μL)
Component C : Assay Buffer 1 bottle (25 mL)
Component D : N-ethylmaleimide standard 1 vial (10 mM, 50 μL )
Component E : DMSO 1 vial (200 μL)
Brief Summary
→ Prepare 20x maleimide reaction mixture (260 μL)
→ Incubate at room temperature for 30 min-1 h
→ Prepare maleimide assay mixture (5 mL total, 50 μL/well)
→ Add maleimide standards or test samples (50 μL)
→ Incubate at room temperature for 5-30 min
→ Read fluorescence at Ex 490 nm/Em 520nm
References
1. Szczepanska A, Espartero JL, Moreno-Vargas AJ, Carmona AT, Robina I, Remmert S, Parish C. (2007) Synthesis
and conformational analysis of novel trimeric maleimide cross-linking reagents. J Org Chem, 72, 6776.
2. Xiao SJ, Wieland M, Brunner S. (2005) Surface reactions of 4-aminothiophenol with heterobifunctional crosslinkers
bearing both succinimidyl ester and maleimide for biomolecular immobilization. J Colloid Interface Sci, 290, 172.
3. Fabisiak JP, Sedlov A, Kagan VE. (2002) Quantification of oxidative/nitrosative modification of CYS(34) in human
serum albumin using a fluorescence-based SDS-PAGE assay. Antioxid Redox Signal, 4, 855.
4. Ghosh SS, Kao PM, McCue AW, Chappelle HL. (1990) Use of maleimide-thiol coupling chemistry for efficient
syntheses of oligonucleotide-enzyme conjugate hybridization probes. Bioconjug Chem, 1, 71.
5. Fujiwara K, Saita T, Kitagawa T. (1988) The use of N-[beta-(4-diazophenyl)ethyl]maleimide as a coupling agent in
the preparation of enzyme-antibody conjugates. J Immunol Methods, 110, 47.
6. Wu CW, Yarbrough LR. (1976) N-(1-pyrene)maleimide: a fluorescent cross-linking reagent. Biochemistry, 15, 2863. |
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Maleimide Quantitation
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Product Code |
Unit |
Price |
Availability |
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5523 |
200 assay |
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