N-Hydroxysulfosuccinimide (sodium salt) (Synonyms: Sulfo-NHS) |
Catalog No.GC44398
One-Click Copy Product Info
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N-Hydroxysulfosuccinimide (sodium) is a non-cleavable ADC linker used in the synthesis of antibody-drug conjugates (ADCs).
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 106627-54-7
Sample solution is provided at 25 µL, 10mM.
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N-Hydroxysulfosuccinimide (sodium salt) is a water-soluble amine-reactive activating reagent. N-Hydroxysulfosuccinimide can be used together with carbodiimides such as 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride. N-Hydroxysulfosuccinimide converts carboxyl groups into more stable sulfo-NHS esters via an O-acylisourea intermediate. The sulfo-NHS esters then form amide bonds with primary amines. The sulfonate group increases aqueous solubility and reduces hydrophobic protein aggregation. N-Hydroxysulfosuccinimide can be used for protein conjugation, antibody biotinylation, cell surface labeling, and carboxylated bead and sensor interface immobilization studies[1-4].
References:[1] Anjaneyulu PS, Staros JV. Reactions of N-hydroxysulfosuccinimide active esters. Int J Pept Protein Res. 1987 Jul;30(1):117-24.
[2] Leichner C, Wulz P, Baus RA, et al. N-Hydroxysulfosuccinimide Esters versus Thiomers: A Comparative Study Regarding Mucoadhesiveness. Mol Pharm. 2019 Mar 4;16(3):1211-1219.
[3] Presentini R. Spectrophotometric method for the quantitative assay of N-hydroxysulfosuccinimide esters including extinction coefficients and reaction kinetics. Anal Biochem. 2017 May 15;525:85-88.
[4] Bogdanov AA Jr, Klibanov AL, Torchilin VP. Protein immobilization on the surface of liposomes via carbodiimide activation in the presence of N-hydroxysulfosuccinimide. FEBS Lett. 1988 Apr 25;231(2):381-4.
An example of magnetic bead-protein conjugation based on N-Hydroxysulfosuccinimide[1-2]
This experimental protocol is compiled from the research content for reference only. Please adjust it according to actual needs.
(1) Reagent preparation: N-Hydroxysulfosuccinimide, EDC.
(2) Reagent setup: ① MES buffer: 100mM MES (pH=4.8); avoid primary amine components like Tris, glycine, BSA. ② Blocking solution: 3M ethanolamine (with 1% BSA, 0.5% Tween-20, pH=8.0) or 1M glycine. ③ Storage solution: PBS (pH=7.4) containing 0.05% Proclin-300.
(3) Magnetic bead activation and coupling: Take appropriate amount of carboxyl magnetic beads (5-10mg/mL), wash twice with MES buffer, perform magnetic separation and discard supernatant. Resuspend the magnetic beads in MES buffer, add EDC and N-Hydroxysulfosuccinimide (recommended final concentration 1.5mg/mL each, or EDC:COOH > 2, NHS:COOH > 20), rotate at room temperature (or 37℃) for activation for 30-60min.
(4) Ligand coupling: After activation, wash the magnetic beads to remove free EDC/N-Hydroxysulfosuccinimide, add protein to be coupled (e.g., SA at 1:25, antibody 20–100μg), adjust volume with MES buffer, rotate at 37℃ for reaction for 2h or react overnight at 4℃. After reaction, perform magnetic separation, add blocking solution, block at 37℃ for 1h (or at room temperature for 30-60min). Wash the magnetic beads 3–5 times with storage solution, resuspend in storage solution, and store at 2–8℃.
Precautions:
(1) N-Hydroxysulfosuccinimide can significantly inhibit O-acylurea hydrolysis and improve coupling yield.
(2) Coupling buffer must not contain primary amine substances to avoid competitive reactions.
(3) For your safety and health, please wear lab coat and disposable gloves during operation.
References:[1] Staros JV, Wright RW, Swingle DM. Enhancement by N-hydroxysulfosuccinimide of water-soluble carbodiimide-mediated coupling reactions. Anal Biochem. 1986 Jul;156(1):220-2.
[2] Staros JV. N-hydroxysulfosuccinimide active esters: bis(N-hydroxysulfosuccinimide) esters of two dicarboxylic acids are hydrophilic, membrane-impermeant, protein cross-linkers. Biochemistry. 1982 Aug 17;21(17):3950-5.
| Cas No. | 106627-54-7 | SDF | |
| Synonyms | Sulfo-NHS | ||
| Canonical SMILES | ON1C(CC(S([O-])(=O)=O)C1=O)=O.[Na+] | ||
| Formula | C4H4NO6S•Na | M.Wt | 217.1 |
| Solubility | PBS (pH 7.2): 10 mg/ml | Storage | Store at -20°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. |
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| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
| Prepare stock solution | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 4.6062 mL | 23.0309 mL | 46.0617 mL |
| 5 mM | 921.2 μL | 4.6062 mL | 9.2123 mL |
| 10 mM | 460.6 μL | 2.3031 mL | 4.6062 mL |
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
g
μL
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 saline, 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.
Quality Control & SDS
- View current batch:
- Purity: >97.00% Appearance: A solid
- COA (Certificate of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 22 reference(s) in Google Scholar.)