A8003-100mg | Sulfo-NHS-LC-Biotin [127062-22-0] Clinisciences

Sulfo-NHS-LC-biotin (Sulfosuccinimidyl 6-(biotinamido) Hexanoate) is an amine-reactive biotinylation agent. As a water-soluble analog of NHS-LC-biotin, Sulfo-NHS-LC-biotin contains a negatively charged sulfonate group on the NHS ring structure, which creates sufficient polarity within the molecule and allows it to be directly added into aqueous reactions without prior dissolution of organic solvents. Sulfo-NHS-LC-biotin covalently binds to amine groups in proteins and other molecules through its NHS ester forming an amide linkage and releasing N-Hydroxysuccinimide. Sulfo-NHS-LC-biotin has a greater length (22.4 A) between a covalently modified molecule and the bicyclic biotin rings than NHS-biotin (13.5 A), which provides the probes a greater opportunity to reach the binding sites deeply buried inside the surface plane of proteins.

Reference

Bioconjugate Techniques , 2nd ed. By Greg T.Hermanson (Pierce Biotechnology, Thermo Fisher Scientific, Rockford, IL). Academic Press (an imprint of Elsevier): London, Amsterdam, Burlington, San Diego . 2008. ISBN 978-0-12-370501-3.

1. Haibo Jin, Chen Yang, et al. "Evaluation of Neurotoxicity in BALB/c Mice following Chronic Exposure to Polystyrene Microplastics." Environ Health Perspect. 2022 Oct;130(10):107002. PMID: 36251724
2. Xihua Yue, Neeraj Tiwari, et al. "Tankyrase-1-mediated degradation of Golgin45 regulates glycosyltransferase trafficking and protein glycosylation in Rab2-GTP-dependent manner." Commun Biol. 2021 Dec 7;4(1):1370. PMID: 34876695
3. Julian Nüchel, Marina Tauber, et al. "An mTORC1-GRASP55 signaling axis controls unconventional secretion to reshape the extracellular proteome upon stress." Mol Cell. 2021 Aug 19;81(16):3275-3293.e12. PMID: 34245671
4. Jia J, Yue X, et al. "KDEL receptor is a cell surface receptor that cycles between the plasma membrane and the Golgi via clathrin-mediated transport carriers." Cell Mol Life Sci. 2020;10.1007/s00018-020-03570-3. PMID: 32562023
5. Wu Z, He D, et al. "A bimodal (SERS and colorimetric) aptasensor for the detection of Pseudomonas aeruginosa." Mikrochim Acta. 2018 Oct 31;185(11):528. PMID: 30382404
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7. Lorin, Charlotte, Isabelle Vögeli, and Ernst Niggli. "Dystrophic cardiomyopathy: role of TRPV2 channels in stretch-induced cell damage." Cardiovascular research 106.1 (2015): 153-162. PMID: 25616416

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• Protein labeling—biotinylate antibodies to facilitate immobilization, purification or detection using streptavidin resins or probes
• Cell surface labeling—do not penetrate the plasma membrane, biotinylates only surface proteins of whole cells
• Amine-reactive—reacts with primary amines (-NH2), such as lysine side-chains, or the N-terminal-amine
• Soluble—charged sulfo-NHS group increases reagent water solubility compared to ordinary NHS-ester compounds
• Irreversible—forms permanent amide bonds; spacer arm cannot be cleaved
• Medium length—spacer arm is 22.4 angstroms; provides excellent balance between adequate length (minimal steric hindrance for biotin binding) and modest mass

Storage	Store at -20°C The product is not stable in solution, please dissolve it immediately before use.
M.Wt	556.6
Cas No.	127062-22-0
Formula	C₂₀H₂₉N₄NaO₉S₂
Synonyms	Biotinamidohexanoic acid 3-sulfo-N-hydroxysuccinimide ester sodium salt
Chemical Name	2,5-dioxo-1-[6-[5-(2-oxo-1,3,3a,4,6,6a-hexahydrothieno[3,4-d]imidazol-4-yl)pentanoylamino]hexanoyloxy]pyrrolidine-3-sulfonic acid;sodium
SDF	Download SDF
Canonical SMILES	C1C(C(=O)N(C1=O)OC(=O)CCCCCNC(=O)CCCCC2C3C(CS2)NC(=O)N3)S(=O)(=O)O.[Na]
Shipping Condition	Small Molecules with Blue Ice, Modified Nucleotides with Dry Ice.
General tips	We do not recommend long-term storage for the solution, please use it up soon.

Biotinylation method [1]:
Sample	Cardiomyocytes
Preparation method	Soluble in Water, DMSO or DMF.
Reaction Conditions	0.5 mg/ml, 37ºC for 2 h
Applications	Sulfo-NHS-LC-Biotin was dissolved in PBS at a concentration of 0.5 mg/ml. then it was incubated with cardiomyocytes at 37ºC for 2 h. After that the cells were washed twice in cold PBS with 100 mM glycine and twice in cold PBS alone to remove the excess biotin reagent. To capture biotinylated proteins, 100 μl of streptavidin agarose resin were incubated with each 100 μg of samples for 16 hours at 4ºC. Captured proteins were eluted at 60ºC for 30 min with Laemmli 2X sample buffer which contain 40 mg/mL of dithiothreitol and they were analyzed by Western blot with primary antibodies.
References: [1]. Charlotte Lorin, Isabelle Vögeli, Ernst Niggli. Dystrophic cardiomyopathy - role of TRPV2 channels in stretch-induced cell damage. Cardiovascular Research, 2015.