ReadiView™ biotin maleimide
Ordering information
Price | |
Catalog Number | |
Unit Size | |
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Additional ordering information
Telephone | 1-800-990-8053 |
Fax | 1-800-609-2943 |
sales@aatbio.com | |
International | See distributors |
Bulk request | Inquire |
Custom size | Inquire |
Shipping | Standard overnight for United States, inquire for international |
Physical properties
Molecular weight | 660.70 |
Solvent | DMSO |
Storage, safety and handling
H-phrase | H303, H313, H333 |
Hazard symbol | XN |
Intended use | Research Use Only (RUO) |
R-phrase | R20, R21, R22 |
Storage | Freeze (< -15 °C) |
UNSPSC | 12352200 |
Alternative formats
ReadiView™ biotin succinimidyl ester |
ReadiView™ biotin amine |
ReadiView™ biotin acid |
ReadiView™ biotin hydrazide |
Related products
Overview | SDSProtocol |
See also: Biotin and Streptavidin
Molecular weight 660.70 |
Biotin/avidin complexes are widely applied for a variety of biological detections. Although a large number of biotin-labeled bioconjugates are commercially available, the accurate determination of biotinylation degree (ratio of biotin/biopolymer) is still a great challenge for biochemists. HABA is still predominantly used for determining the degree of biotinylation (through its absorption with the extinction coefficient = 34,000/M-1cm-1). When a biotin-containing sample is added, the biotin binds strongly to avidin and displaces the weakly bound HABA. The resulting decrease in absorbance relates to the amount of biotin. However there are many factors that affect the accuracy of HABA method, making this method unreliable for many biotin-labeled conjugates. Our ReadiView™ biotin contains specially designed Color Tag (CT) that makes the biotinylation degree readily accessible by simply calculating the corrected absorption ratio of 280 nm/385 nm. Our specially designed tag has very minimal effect on the biotin binding affinity, and its absorption maximum was designed to make the tag have minimal quenching effect on the most fluorophores that are used for labeling avidins.
Example protocol
SAMPLE EXPERIMENTAL PROTOCOL
Labeling Proteins with ReadiView™ biotin maleimide
- 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 ReadiView™ biotin maleimide in DMSO at a concentration of 5 - 10 mg/mL.
- Mix the ReadiView™ biotin 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 ReadiView™ biotin maleimide
- Dissolve ReadiView™ biotin 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 ReadiView™ biotin 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 | 151.355 µL | 756.773 µL | 1.514 mL | 7.568 mL | 15.135 mL |
5 mM | 30.271 µL | 151.355 µL | 302.709 µL | 1.514 mL | 3.027 mL |
10 mM | 15.135 µL | 75.677 µL | 151.355 µL | 756.773 µL | 1.514 mL |
Molarity calculator
Enter any two values (mass, volume, concentration) to calculate the third.
Mass (Calculate) | Molecular weight | Volume (Calculate) | Concentration (Calculate) | Moles | ||||
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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
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
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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
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Journal: J Comp Neurol (2004): 86
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
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
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
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
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
Authors: Xu H, Zhang S, Liu D, Liang CC.
Journal: Mol Biotechnol (2001): 183