35013-72-0

Basic information

• Product Name: BIOTIN-NHS
• Synonyms: SUCCINIMIDO-(+)-BIOTIN;SUCCINIMIDYL D-BIOTIN;N-HYDROXYSUCCINIMIDOBIOTIN;NHS-BIOTIN;NHS-D-BIOTIN;N-SUCCINIMIDYL D-BIOTINATE;N-SUCCINIMIDO-(+)-BIOTIN;5-(2-OXO-HEXAHYDRO-THIENO[3,4-D]IMIDAZOL-6-YL)-PENTANOIC ACID 2,5-DIOXO-PYRROLIDIN-1-YL ESTER
• CAS NO: 35013-72-0
• Molecular Formula: C₁₄H₁₉N₃O₅S
• Molecular Weight: 341.38
• EINECS: 1592732-453-0
• Product Categories: Biotin Derivatives;Cross Linking Reagents;BiotinylationReagents;Biotinylation Reagents;biotinyl
• Mol File: 35013-72-0.mol

Chemical Properties

• Melting Point: 212-214 °C
• Appearance: White to off-white/powder
• Density: 1.45 g/cm³
• Refractive Index: 1.616
• Storage Temp.: −20°C
• Solubility: DMF: ≤50 mg/mL Stable for at least one month in dry D
• PKA: 13.89±0.40(Predicted)
• Stability: Stable. Incompatible with strong oxidizing agents.
• BRN: 4720781
• CAS DataBase Reference: BIOTIN-NHS(CAS DataBase Reference)
• NIST Chemistry Reference: BIOTIN-NHS(35013-72-0)
• EPA Substance Registry System: BIOTIN-NHS(35013-72-0)

Safety Data

• Safety Statements: 22-24/25
• WGK Germany: 3
• F: 10-23
• Hazardous Substances Data: 35013-72-0(Hazardous Substances Data)

Usage

Uses

Used in Biochemical Research:
BIOTIN-NHS is used as an amine reactive biotinylation reagent for the biotinylation of proteins and peptides. It couples with primary amines in the pH range of 6.5-8.5, making it an essential tool in various biochemical research applications.
Used in Diagnostics and Detection:
In the diagnostics industry, BIOTIN-NHS is used as a biotinylation agent to create biotinylated probes, which can be used for the detection and identification of specific target molecules in various diagnostic assays.
Used in Drug Delivery Systems:
BIOTIN-NHS can be employed in the development of targeted drug delivery systems, where biotinylated molecules can be used to specifically bind to receptors or other molecular targets on the surface of cells, enhancing the selectivity and efficacy of drug delivery.
Used in Surface Functionalization:
In the materials science and engineering industry, BIOTIN-NHS is used as an amino reactive biotin reagent for preparing biotinylated surfaces. These surfaces can be utilized in various applications, such as biosensors, cell culture substrates, and immobilization of biomolecules for analytical purposes.
Used in Immunoassays:
In the biotechnology industry, BIOTIN-NHS is used as a key component in the development of immunoassays, where biotinylated antibodies or other binding molecules can be employed for the detection and quantification of specific analytes.

Purification Methods

Recrystallise the ester from refluxing isoPrOH and dry it in a vacuum over P2O5 + KOH. [Jasiewicz et al. Exp Cell Biol 100 213 1976.]

InChI:InChI=1/C14H19N3O5S/c18-10-5-6-11(19)17(10)22-12(20)4-2-1-3-9-13-8(7-23-9)15-14(21)16-13/h8-9,13H,1-7H2,(H2,15,16,21)/t8-,9-,13-/m0/s1

Upstream product

• 6066-82-6 1-hydroxy-pyrrolidine-2,5-dione 
• 101187-31-9 (3aS,4S,6aR)-4-[5-(1H-imidazol-1-yl)-5-oxopentyl]tetrahydro-1H-thieno-[3,4-d]imidazol-2(3H)-one 
• 74124-79-1 di(succinimido) carbonate 
• 58-85-5 biotin 
• 105832-38-0 O-(N-succinimidyl)-N,N,N',N'-tetramethyluronium tetrafluoroborate 

Downstream Products

• 153712-60-8 5-((3aR,6S,6aS)-2-Oxo-hexahydro-thieno[3,4-d]imidazol-6-yl)-pentanoic acid [2-(4-hydroxy-3,5-diiodo-phenyl)-ethyl]-amide 
• 112242-36-1 C₁₈H₂₄⁽¹²⁵⁾IN₃O₃S 
• 62062-43-5 Boc-Lys(biotinyl)-OH 
• 175885-18-4 tert-butyl-N-(2-(2-(2-(5-(2-oxo-1,3,3a,4,6,6a-hexahydrothieno(3,4-d)imidazol-6-yl)pentanoylamino)ethoxy)ethoxy)ethyl)carbamate 
• 184296-68-2 (2R,4S,5R)-5-[Bis-(4-methoxy-phenyl)-phenyl-methoxymethyl]-4-hydroxy-tetrahydro-furan-2-carboxylic acid {2-[5-((3aR,6S,6aS)-2-oxo-hexahydro-thieno[3,4-d]imidazol-6-yl)-pentanoylamino]-ethyl}-amide 
• 188584-23-8 5-((3aR,6S,6aS)-2-Oxo-hexahydro-thieno[3,4-d]imidazol-6-yl)-pentanoic acid undec-10-ynylamide 
• 188257-55-8 N-biotinyl-(4,4'-dimethoxytrityloxymethyl)-5-(hydroxymethyl)benzylamine 
• 402725-75-1 12-benzylthiododecanoic-(8-biotinoylamido-3,6-dioxaoctyl)amide 
• 440359-01-3 N-[3-(N-cholesteryl-2-nitrobenzenesulfonamido)propyl]biotinamide 
• 157720-52-0 N-(4-nitrobenzyl)biotin amide 
• 918300-29-5 diethyl 2,2’-((5-((2-(5-((3aS,4S,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamido)ethyl)carbamoyl)-1,3-phenylene)bis(oxy))diacetate 
• 773888-45-2 N-(prop-2-ynyl)biotinamide 
• 908007-17-0 N-(3-azidopropyl)-5-((3aS,4S,6aR)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamide 
• 576-19-2 biocytin 

Relevant articles and documents

Synthesis and properties of a biotin-tagged NHC-gold complex

The first synthesis of a biotin-tagged NHC-gold complex is described. The key step is the coupling of alkyne-substituted biotin derivative 4b with azido-imidazolium salt 8a by copper-catalyzed azide alkyne cycloaddition (CuAAC). Gold complex 2 catalyzes the cycloisomerization of α-hydroxyallenes to 2,5-dihydrofurans.

Synthesis and Biological Actions of HIghly Potent and Prolonged Acting Biotin-Labeled Melanotropins

Biocytin derivatives of a superpotent analogue of α-melanotropin, 4,D-Phe7>-α-MSH, were prepared. α-Bct-Ser1,Nle4,D-Phe7>-α-MSH and α-dodecanoyl-Ser1,Nle4,D-Phe7>-α-MSH were synthesized by solid-phase techniques, and the coupling of biotin and 12-aminododecanoic acid was achieved through their succinimido esters.These melanotropins possessed almost inentical actions to 4,D-Phe7>-α-MSH as determined by several melanocyte bioassays.Both biocytin derivatives were highly potent agonists and exhibited prolonged biological activity as determined in the frog and lizard skin bioassays.Both biotinylated peptides were at least equipotent to α-MSH in stimulating Cloudman S91 mouse melanoma tyrosinase activity.The analogues were resistant to inactivation by α-chymotrypsin.

Synthesis and Avidin Binding of Ruthenium Complexes Functionalized with a Light-Cleavable Free Biotin Moiety

In this work the synthesis, photochemistry, and streptavidin interaction of new [Ru(tpy)(bpy)(SRR′)](PF6)2 complexes where the R′ group contains a free biotin ligand, are described. Two different ligands SRR′ were investigated: An asymmetric ligand 1 where the Ru-bound thioether is a N-acetylmethionine moiety linked to the free biotin fragment via a triethylene glycol spacer and a symmetrical ligand 2 containing two identical biotin moieties. The coordination of these two ligands to the precursor [Ru(tpy)(bpy)Cl]Cl was studied in water at 80 °C. In such conditions the coordination of the asymmetric ligand 1 occurred under thermodynamic control. After the reaction, a mononuclear and a binuclear complex were isolated. In the mononuclear complex, the ratio of methionine- {[6](PF6)2} vs. biotin-bound {[7](PF6)2} regioisomer was 5.3 and the free biotin fragment of [6](PF6)2 allowed to purify it from its isomer [7](PF6)2 at small scales using avidin affinity chromatography. Coordination of the symmetrical ligand 2 afforded [Ru(tpy)(bpy)(2)](PF6)2 {[8](PF6)2} in synthetically useful scales (100 mg), good yield (82 %), and without traces of the binuclear impurity. In this complex, one of the biotin remains free whereas the second one is coordinated to ruthenium. Photochemical release of ligand 2 from [8](PF6)2 occurred upon blue light irradiation (465 nm) with a photosubstitution quantum yield of 0.011 that was independent of the binding of streptavidin to the free biotin ligand.

Synthesis of biotin labelled cap analogue - Incorporable into mRNA transcripts and promoting cap-dependent translation

Analogues of the eukaryotic messenger RNA 5′ end (m7G cap) are useful tools for studying mRNA fate and serve as reagents for in vitro preparation of 5′ capped mRNAs. We designed a biotin-labeled dinucleotide cap analogue that can be incorporated into transcripts to produce 5′-capped and biotinylated mRNAs which retain their biological functionality and may be employed for biotin-(strept)avidin technologies.

mRNA Cap Modification through Carbamate Chemistry: Synthesis of Amino- and Carboxy-Functionalised Cap Analogues Suitable for Labelling and Bioconjugation

A simple and efficient synthesis of dinucleotide mRNA 5′ cap analogues functionalised by the addition of an amine or carboxylic acid group onto one of the ribose rings is reported. Selection of the modification sites was dictated by the recognition specificity for the 5′ cap by various cap-binding proteins, including eIF4E, DcpS, and Dcp2, as well as by bacteriophage RNA polymerase, which enables the incorporation of the synthetic cap into the mRNA chain. Some analogues were further modified by O-to-CH2 substitution in the triphosphate bridge to confer resistance to specific hydrolases. Spatial crowding and structural rigidity were balanced by use of diamines and amino acids of different lengths attached to the ribose through a carbamate moiety. The impact of these substitutions on the conformational equilibria of the ribose components was investigated by 1H NMR spectroscopy. The utility of the analogues for labelling with fluorescent dyes and affinity tags was demonstrated with the aid of NHS activation chemistry. A series of amino- and carboxy-functionalised dinucleotide 5′ cap analogues have been synthesised by use of carbamate chemistry. The compounds are suitable for labelling with biotin or fluorescent tags by employment of an in situ NHS activation strategy,

Synthesis of Na2S2O4 mediated cleavable affinity tag for labeling of O-GlcNAc modified proteins via azide-alkyne cycloaddition

A facile and convergent procedure for the synthesis of azobenzene-based probe was reported, which could selectively release interested proteins conducted with sodium dithionite. Besides, the cleavage efficiency is closely associated with the structural features, in which an ortho-hydroxyl substituent is necessary for reactivity. In addition, the azobenzene tag applied in the Ac4GlcNAz-labled proteins demonstrated high efficiency and selectivity in comparison with Biotin-PEG4-Alkyne, which provides a useful platform for enrichment of any desired bioorthogonal proteomics.

LIGAND LINKER SUBSTRATE

Ligand functionalized substrate including a solid substrate, which has been modified to provide grafted catching ligand groups covalently bound via a linker, methods of preparing said ligand functionalized substrate and the use thereof, such as to increase binding rate and the dynamic binding capacity (DBC).

Design and synthesis of biotinylated cardiac glycosides for probing Nur77 protein inducting pathway

The orphan nuclear receptor Nur77 (also known as TR3 or nerve growth factor-induced clone B NGFI-B) functions as a nuclear transcription factor in the regulation of target gene expression and plays a critical role in the regulation of differentiation, proliferation, apoptosis, and survival of many different cell types. Recent studies demonstrate that Nur77 also involves many important physiological and pathological processes including cancer, inflammation and immunity, cardiovascular diseases, and bone diseases. Our previous studies showed that cardiac glycosides could induce the expression of Nur77 protein and its translocation from the nucleus to the cytoplasm and subsequent targeting to mitochondria, leading to apoptosis of cancer cells. In order to probe the Nur77 protein inducting pathway, we designed and synthesized a series of novel biotinylated cardiac glycosides from β-Antiarin and α-Antiarin, two typical cardiac glycosides from the plant of Antiaris toxicaria. The induction of Nur77 protein expression of these biotinylated cardiac glycosides and their inhibitory effects on NIH-H460 cancer cell proliferation were evaluated. Results displayed that some biotinylated cardiac glycosides could significantly induce the expression of Nur77 protein comparable with their parent compounds β-Antiarin and α-Antiarin. Also, their streptavidin binding activities were evaluated. Among them, biotinylated cardiac glycosides P4b and P5a exhibited significant effect on the induction of Nur77 expression along with high binding capacity with streptavidin, suggesting that they can be used as probes for probing Nur77 protein inducting pathway.

High-Throughput Chemical Probing of Full-Length Protein-Protein Interactions

Human biology is regulated by a complex network of protein-protein interactions (PPIs), and disruption of this network has been implicated in many diseases. However, the targeting of PPIs remains a challenging area for chemical probe and drug discovery. Although many methodologies have been put forth to facilitate these efforts, new technologies are still needed. Current biochemical assays for PPIs are typically limited to motif-domain and domain-domain interactions, and assays that will enable the screening of full-length protein systems, which are more biologically relevant, are sparse. To overcome this barrier, we have developed a new assay technology, PPI catalytic enzyme-linked click chemistry assay or PPI cat-ELCCA, which utilizes click chemistry to afford catalytic signal amplification. To validate this approach, we have applied PPI cat-ELCCA to the eIF4E-4E-BP1 and eIF4E-eIF4G PPIs, key regulators of cap-dependent mRNA translation. Using these examples, we have demonstrated that PPI cat-ELCCA is amenable to full-length proteins, large (>200 kDa) and small (～12 kDa), and is readily adaptable to automated high-throughput screening. Thus, PPI cat-ELCCA represents a powerful new tool in the toolbox of assays available to scientists interested in the targeting of disease-relevant PPIs.

BIOCONJUGATES OF HETEROCYCLIC COMPOUNDS

The invention provides bioconjugates of heterocylic compounds such as S-adenosylmethionine and S-adenosylhomocysteine with biotin or digoxigenin. The bioconjugates also include carbon and nitrogen linker moiteies of varying length that are used to attach such compounds to biotin or digoxigenin. The conjugates are useful in immunoassays. The invention provides a method for detecting SAM and SAH, comprising the steps of: (a) preparing the following components: (i) bio-conjugates of SAM, SAM analogs or SAH; (ii) an europium, a terbium cryptate or other fluorophore as a donor that has a specific ligand for the tracer in the bio-conjugates of (i); (iii) an acceptor fluorescent dye that has the excitation spectra overlap those of donor's emissions and has an antibody specific for SAM or SAH labeled; (b) addition of the biological fluid containing said SAM or SAH; and (c) spectroscopic measurement of the fluorescence of the donor and the fluorescence of from the acceptor.