Biotin (Synonyms: D-Biotin, NSC 63865, Vitamin B7) |
Catalog No.GC11194 |
A coenzyme of carboxylase enzymes
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 58-85-5
Sample solution is provided at 25 µL, 10mM.
Quality Control & SDS
- View current batch:
- Purity: >99.50%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Biotinylation method [1]: | |
Sample |
Proteins |
Preparation method |
Soluble in DMSO > 10 mM. General tips for obtaining a higher concentration: Please warm the tube at 37 ℃ for 10 minutes and/or shake it in the ultrasonic bath for a while. Stock solution can be stored below -20℃ for several months. |
Reaction Conditions |
room temperature for 1 h |
Applications |
Biotin-labeling of proteins: Biotinyl-N-hydroxysuccinimide (BNHS) was used for introducing biotin moieties into proteins. In order to obtain the desired molar ratio of BNHS to free amino groups of the protein, a solution of 0.1 M NaHCO: containing 10 mg/ml of protein was mixed with various volumes of 0.1 M solution of BNHS. The reaction mixture was incubated at room temperature for 1 h and then dialyzed for 24 h at 4℃ against several changes of PBS. After dialysis, an equal volume of glycerol was added and the preparation was kept at -20℃ until used. A fresh solution of BNHS was prepared each time immediately before use. The 2, 4, 6-trinitrobenzene sulfonic acid procedure was used to determine the free amino groups in native and biotin-substituted proteins. Total biotin covalently bound to IgG was determined by an avidin-binding assay. |
References: [1]. JEAN-LUC GUESDON, THERESE TERNYNCK AND STRATIS AVRAMEAS. The Use of Avidin-Biotin Interaction in Immunoenzymatic Techniques. THE JOURNAL OF HISTOCHEMISTRY AND CYTOCHEMISTRY. 1979. |
NHS-biotin modification as a specific lysine probe coupled to mass spectrometry detection is increasingly used over the past years for assessing amino acid accessibility of proteins or complexes as an alternative when well-established methods are challenged1.
Numerous applications have already been reported2, with increasing use of N-HydroxySuccinimide- biotin (NHS-biotin) as a specific lysine probe3. As an example, determination of surface accessibility of amino acids may be useful to delineate protein–protein interfaces as shown in epitopemapping4.
Labeling of amino acids containing hydroxyl groups has been already observed for the Sulfo-NHS-biotin reagent reacting on model peptides33 and for 3,30-Dithiobis[sulfosuccinimidyl propionate] (DTSSP)5.It has been shown that serine, tyrosine, and threonine residues readily react with Sulfo- NHS-biotin if the hydroxyl-containing group is located two positions from a histidine residue6. It was proposed that the enhanced intrinsic reactivity of these hydroxyl groups was due to an interaction with the histidyl residue via hydrogen bonding that contributes to increase their nucleophilicity.
The pioneering observationsmade with DTSSP are also detected with NHS-biotin derivatives: hydroxyl groups of serine and tyrosine residues also reactwith primary amine reagents and serine label may be lost resulting in a dehydrated peptide. Use of biotin reagents allows affinity enrichment by means of streptavidin3, as well as absolute quantitation of the labels introduced per polypeptide by means of a spectrophotometric assay.
Use of the complete set of Sulfo-NHS-biotin reagents allows an efficient and reliable assignment of the different ions detected.
Reference:
1. G. Gabant, J. Augier et al, Assessment of solvent residues accessibility using three Sulfo-NHS-biotin reagents in parallel: application to footprint changes of a methyltransferase upon binding its substrate, J. Mass Spectrom. 2008; 43: 360–370
2. Glocker MO, Borchers C, Fiedler W, Suckau D, Przybylski M. Molecular characterization of surface topology in protein tertiary structures by amino-acylation and mass spectrometric peptide mapping. Bioconjugate Chem. 1994; 5: 583.
3. Azim-Zadeh O, Hillebrecht A, Linne U, Marahiel MA, Klebe G, Lingelbach K, Nyalwidhe J. Use of biotin derivatives to probe conformational changes in proteins. Journal of Biological Chemistry 2007; 282: 21609
4. Borch J, Jorgensen TJ, Roepstorff P. Mass spectrometric analysis of protein interactions. Current Opinion in Chemical Biology2005; 9: 509.
5. Swaim CL, Smith JB, Smith DL. Unexpected products from the reaction of the synthetic cross-linker 3, 3' dithiobis(sulfosuccinimidyl propionate), DTSSP with peptides. Journal of the American Society for Mass Spectrometry 2004; 15:736.
6. Miller BT, Kurosky A. Elevated intrinsic reactivity of seryl hydroxyl groups within the linear peptide triads His-Xaa-Ser or Ser-Xaa-His. Biochemical and Biophysical Research Communications 1993; 196: 461.
Cas No. | 58-85-5 | SDF | |
Synonyms | D-Biotin, NSC 63865, Vitamin B7 | ||
Chemical Name | 5-[(3aS,4S,6aR)-2-oxo-1,3,3a,4,6,6a-hexahydrothieno[3,4-d]imidazol-4-yl]pentanoic acid | ||
Canonical SMILES | C1C2C(C(S1)CCCCC(=O)O)NC(=O)N2 | ||
Formula | C10H16N2O3S | M.Wt | 244.31 |
Solubility | ≥ 24.4mg/mL in DMSO | Storage | Store at 2-8°C |
General tips | Please select the appropriate solvent to prepare the stock solution according to the
solubility of the product in different solvents; once the solution is prepared, please store it in
separate packages to avoid product failure caused by repeated freezing and thawing.Storage method
and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored
at -20°C, please use it within 1 month. To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time. |
||
Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. |
Prepare stock solution | |||
1 mg | 5 mg | 10 mg | |
1 mM | 4.0932 mL | 20.4658 mL | 40.9316 mL |
5 mM | 0.8186 mL | 4.0932 mL | 8.1863 mL |
10 mM | 0.4093 mL | 2.0466 mL | 4.0932 mL |
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)
Calculation results:
Working concentration: mg/ml;
Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )
Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.
Average Rating: 5
(Based on Reviews and 30 reference(s) in Google Scholar.)GLPBIO products are for RESEARCH USE ONLY. Please make sure your review or question is research based.
Required fields are marked with *