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Biotin C2 maleimide
Biotin and biotin derivatives bind with high affinity to both avidins and streptavidins. Biotin and its derivatives can be conjugated to many biomolecules without significantly altering the biological activity of the target molecules since biotin is relatively a small molecule. A biopolymer (such as proteins) can react with several molecules of biotin that, in turn, can each bind one avidin. This characteristic greatly increases the sensitivity of many biological assays. Biotin derivatives are widely used for biological detections and purification. This biotin derivative is used to modify thiol group.
Example protocol
SAMPLE EXPERIMENTAL PROTOCOL
Labeling Proteins with Biotin C2 Malemide
- Dissolve your thiol-containing protein at concentration of 1 - 10 mg/mL (3 - 10 mg is the optimal labeling concentration) using PBS buffer (20 mM, pH 7.2).
- Dissolve the Biotin C2 Maleimide in DMSO at a concentration of 5 - 10 mg/mL.
- Mix the Biotin C2 Maleimide and protein solution at 2:1 molar ratio of biotin/protein, and shake the reaction mixture at room temperature for 2 - 4 hours.
- Filter the reaction mixture through a protein spin column for 100 µg to 1 mg protein labeling reaction. If the reaction scale is larger than 1 mg, purify the conjugate using gel filtration on a properly sized Sephadex G-25 column.
- Collect the desired fractions for your immediate use or freeze dry them for future use.
Labeling Small Molecules with Biotin C2 Malemide
- Dissolve Biotin C2 Maleimide (10 - 15 mg/mL) and your thiol-containing molecule in DMSO at 1:1.2 molar ratio of biotin/thiol-containing molecule.
- Stir the reaction mixture at room temperature for 2 - 4 hours.
- Purify the conjugate using HPLC (ammonium acetate/water and acetonitrile, pH 7.0).
- Collect and pool the desired fractions.
- Combine and freeze-dry the pooled fractions.
Calculators
Common stock solution preparation
Table 1. Volume of DMSO needed to reconstitute specific mass of Biotin C2 maleimide to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.
| 0.1 mg | 0.5 mg | 1 mg | 5 mg | 10 mg | |
| 1 mM | 272.896 µL | 1.364 mL | 2.729 mL | 13.645 mL | 27.29 mL |
| 5 mM | 54.579 µL | 272.896 µL | 545.792 µL | 2.729 mL | 5.458 mL |
| 10 mM | 27.29 µL | 136.448 µL | 272.896 µL | 1.364 mL | 2.729 mL |
Molarity calculator
Enter any two values (mass, volume, concentration) to calculate the third.
| Mass (Calculate) | Molecular weight | Volume (Calculate) | Concentration (Calculate) | Moles | ||||
| / | = | x | = |
Product family
| Name | Correction Factor (280 nm) |
| AMCA C2 Maleimide | 0.153 |
| EDANS C2 maleimide | 0.107 |
| DABCYL C2 maleimide | 0.516 |
| XFD647 C2 Maleimide *Same Structure to Alexa Fluor™ 647 C2 Maleimide* | 0.03 |
Citations
View all 3 citations: Citation Explorer
A low-cost, high-performance system for fluorescence lateral flow assays
Authors: Lee, Linda G and Nordman, Eric S and Johnson, Martin D and Oldham, Mark F
Journal: Biosensors (2013): 360--373
Authors: Lee, Linda G and Nordman, Eric S and Johnson, Martin D and Oldham, Mark F
Journal: Biosensors (2013): 360--373
Implementation of P22 viral capsids as nanoplatforms
Authors: Kang, Sebyung and Uchida, Masaki and O'Neil, Alison and Li, Rui and Prevelige, Peter E and Douglas, Trevor
Journal: Biomacromolecules (2010): 2804--2809
Authors: Kang, Sebyung and Uchida, Masaki and O'Neil, Alison and Li, Rui and Prevelige, Peter E and Douglas, Trevor
Journal: Biomacromolecules (2010): 2804--2809
Calcium-dependent activation of transglutaminase 2 by nanosecond pulsed electric fields
Authors: Morotomi-Yano, Keiko and Yano, Ken-ichi
Journal: FEBS Open Bio
Authors: Morotomi-Yano, Keiko and Yano, Ken-ichi
Journal: FEBS Open Bio
References
View all 78 references: Citation Explorer
Evaluation of a new biotin-DOTA conjugate for pretargeted antibody-guided radioimmunotherapy (PAGRIT((R)))
Authors: Urbano N, Papi S, Ginanneschi M, De Santis R, Pace S, Lindstedt R, Ferrari L, Choi S, Paganelli G, Chinol M.
Journal: Eur J Nucl Med Mol Imaging. (2006)
Authors: Urbano N, Papi S, Ginanneschi M, De Santis R, Pace S, Lindstedt R, Ferrari L, Choi S, Paganelli G, Chinol M.
Journal: Eur J Nucl Med Mol Imaging. (2006)
A biotin-protein bond with stability in plasma
Authors: Bogusiewicz A, Mock NI, Mock DM.
Journal: Anal Biochem (2005): 98
Authors: Bogusiewicz A, Mock NI, Mock DM.
Journal: Anal Biochem (2005): 98
Instability of the biotin-protein bond in human plasma
Authors: Bogusiewicz A, Mock NI, Mock DM.
Journal: Anal Biochem (2004): 156
Authors: Bogusiewicz A, Mock NI, Mock DM.
Journal: Anal Biochem (2004): 156
Design, synthesis, and evaluation of 5'-S-aminoethyl-N(6)- azidobenzyl-5'-thioadenosine biotin conjugate: a bifunctional photoaffinity probe for the es nucleoside transporter
Authors: Addo JK, Buolamwini JK.
Journal: Bioconjug Chem (2004): 536
Authors: Addo JK, Buolamwini JK.
Journal: Bioconjug Chem (2004): 536
Biotin is endogenously expressed in select regions of the rat central nervous system
Authors: McKay BE, Molineux ML, Turner RW.
Journal: J Comp Neurol (2004): 86
Authors: McKay BE, Molineux ML, Turner RW.
Journal: J Comp Neurol (2004): 86
Page updated on November 20, 2024
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