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Biotin C2 maleimide

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Fax1-800-609-2943
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Physical properties
Molecular weight366.44
SolventDMSO
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
StorageFreeze (< -15 °C)
UNSPSC12352200
Alternative formats
Biotin C2 Azide
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OverviewpdfSDSpdfProtocol


CAS
139554-72-6
Molecular weight
366.44
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
  1. 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).
  2. Dissolve the Biotin C2 Maleimide in DMSO at a concentration of 5 - 10 mg/mL.
  3. 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.
  4. 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.
  5. Collect the desired fractions for your immediate use or freeze dry them for future use. 

Labeling Small Molecules with Biotin C2 Malemide
  1. 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.
  2. Stir the reaction mixture at room temperature for 2 - 4 hours.
  3. Purify the conjugate using HPLC (ammonium acetate/water and acetonitrile, pH 7.0).
  4. Collect and pool the desired fractions.
  5. 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 mg0.5 mg1 mg5 mg10 mg
1 mM272.896 µL1.364 mL2.729 mL13.645 mL27.29 mL
5 mM54.579 µL272.896 µL545.792 µL2.729 mL5.458 mL
10 mM27.29 µL136.448 µL272.896 µL1.364 mL2.729 mL

Molarity calculator

Enter any two values (mass, volume, concentration) to calculate the third.

Mass (Calculate)Molecular weightVolume (Calculate)Concentration (Calculate)Moles
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Product Family


NameCorrection Factor (280 nm)
AMCA C2 Maleimide0.153
EDANS C2 maleimide0.107
DABCYL C2 maleimide0.516

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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
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
Calcium-dependent activation of transglutaminase 2 by nanosecond pulsed electric fields
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)
A biotin-protein bond with stability in plasma
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
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
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
Probing the interaction of the biotin-avidin complex with the relaxivity of biotinylated Gd-DTPA
Authors: Langereis S, Kooistra HA, van Genderen MH, Meijer EW.
Journal: Org Biomol Chem (2004): 1271
Immunoassay of total prostate-specific antigen using europium(III) nanoparticle labels and streptavidin-biotin technology
Authors: Huhtinen P, Soukka T, Lovgren T, Harma H.
Journal: J Immunol Methods (2004): 111
Volume of RBCs, 24- and 48-hour posttransfusion survivals, and the lifespan of (51)Cr and biotin-X-N-hydroxysuccinimide (NHS)-labeled autologous baboon RBCs: effect of the anticoagulant and blood pH on (51)Cr and biotin-X-NHS elution in vivo
Authors: Valeri CR, Pivacek LE, Cassidy GP, Ragno G.
Journal: Transfusion (2002): 343
Endogenous biotin staining in a subset of spinal neuronal cell bodies: a potential confounding factor for neuroanatomical studies
Authors: Berkowitz A., undefined
Journal: Brain Res (2002): 98
End-labeling of long DNA fragments with biotin and detection of DNA immobilized on magnetic beads
Authors: Xu H, Zhang S, Liu D, Liang CC.
Journal: Mol Biotechnol (2001): 183