2-Nitro-5-thiocyanatobenzoic Acid (Synonyms: NTCB) |
رقم الكتالوجGC42185
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2-Nitro-5-thiocyanatobenzoic acid (NTCB) هو كاشف شديد التفاعل ينقل مجموعة cyano الخاصة به بسرعة إلى ثيولات نيوكليوفيليك
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 30211-77-9
Sample solution is provided at 25 µL, 10mM.
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2-Nitro-5-thiocyanatobenzoic Acid is a highly reactive cyano group donor. 2-Nitro-5-thiocyanatobenzoic Acid can rapidly transfer its cyano group to a nucleophilic thioester, converting the cysteine thiol groups in proteins into S-cyano derivatives, while simultaneously achieving functionalization modification at the C-terminus of proteins through the formation of a 1-acyl-2-iminothiazolidine intermediate. 2-Nitro-5-thiocyanatobenzoic Acid can be used in research related to protein chemical synthesis, cysteine-specific cleavage and labeling, and polypeptide drug development[1-4].
References:
[1] Tang HY, Speicher DW. Identification of alternative products and optimization of 2-nitro-5-thiocyanatobenzoic acid cyanylation and cleavage at cysteine residues. Anal Biochem. 2004 Nov 1;334(1):48-61.
[2] Belghazi M, Klett D, Cahoreau C, et al. Nitro-thiocyanobenzoic acid (NTCB) reactivity of cysteines beta100 and beta110 in porcine luteinizing hormone: metastability and hypothetical isomerization of the two disulfide bridges of its beta-subunit seatbelt. Mol Cell Endocrinol. 2006 Mar 9;247(1-2):175-82.
[3] Koehn H, Clerens S, Deb-Choudhury S, et al. Higher sequence coverage and improved confidence in the identification of cysteine-rich proteins from the wool cuticle using combined chemical and enzymatic digestion. J Proteomics. 2009 Dec 1;73(2):323-30.
[4] Chu Y, Lee EY, Reimann EM, et al. Effect of activation of protein phosphatase 1 on sulfhydryl reactivity. Arch Biochem Biophys. 1996 Oct 1;334(1):83-8.
2-Nitro-5-thiocyanatobenzoic Acid-Induced DNase Inactivation[1]
This protocol is adapted from research data and provided for reference only. Adjustments may be necessary based on specific experimental requirements.
1. Reagent Preparation: Dissolve 2-Nitro-5-thiocyanatobenzoic Acid in an appropriate buffer as a working solution (33mM), including Iodoacetate (67mM), N-ethylmorpholine acetate (0.2mM); prepare DNase I (10 mg).
2. DNase Treatment and Inactivation with 2-Nitro-5-thiocyanatobenzoic Acid: In the presence of Ca²⁺, incubate DNase I with the 2-Nitro-5-thiocyanatobenzoic Acid working solution at room temperature. Monitor the inactivation kinetics by periodically measuring the remaining DNase activity. Complete inactivation typically takes several hours.
3. Peptide Bond Cleavage Analysis and Modification Site Identification: After treatment with 2-Nitro-5-thiocyanatobenzoic Acid, peptide bond cleavage occurs at the N-terminus of specific serine (Ser) and threonine (Thr) residues in the enzyme molecule. Separate the resulting polypeptide fragments by gel chromatography and SDS-polyacrylamide gel electrophoresis. Analyze the amino acid composition by eluting protein bands from the gel.
Precautions:
(1) Ca²⁺ is an essential cofactor for DNase inactivation induced by 2-Nitro-5-thiocyanatobenzoic Acid.
(2) For your safety and health, please wear a lab coat and disposable gloves during operation.
References:
[1] Liao TH, Wadano A. Inactivation of DNase by 2-nitro-5-thiocyanobenzoic acid. II. Serine and threonine are the sites of reaction on the DNase molecule. J Biol Chem. 1979 Oct 10;254(19):9602-7.
| Cas No. | 30211-77-9 | SDF | |
| المرادفات | NTCB | ||
| Canonical SMILES | OC(C1=C([N+]([O-])=O)C=CC(SC#N)=C1)=O | ||
| Formula | C8H4N2O4S | M.Wt | 224.2 |
| الذوبان | DMF: 25 mg/ml,DMSO: 11 mg/ml,Ethanol: 25 mg/ml,PBS (pH 7.2): 0.3 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. |
||
| 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 | 4.4603 mL | 22.3015 mL | 44.603 mL |
| 5 mM | 892.1 μL | 4.4603 mL | 8.9206 mL |
| 10 mM | 446 μL | 2.2302 mL | 4.4603 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 6 reference(s) in Google Scholar.)