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| Targets |
The primary molecular target of NSP-SA-NHS is not a biological receptor or enzyme, but rather the antigen, antibody, protein, or nucleic acid to which it is conjugated for detection purposes. As a chemiluminescent labeling reagent, NSP-SA-NHS is used to label biomolecules for use in immunoassays and other detection systems. The NHS (N-hydroxysuccinimide) ester group reacts with primary amines on proteins or other biomolecules to form stable amide bonds, covalently attaching the acridinium ester label to the target molecule. The labeled biomolecule can then be used in chemiluminescent immunoassays for the detection of analytes.
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| ln Vitro |
NSP-SA-NHS labeling [1] (1) 0.2 mg of NSP-SA-NHS and 1 mg of BSA are dissolved in 40 uL DMF and 0.3 mL PBS, respectively (solution A and B). (3) Use 500 mL of additional water and sterilize at 4°C for 2 days, changing the supplementary water (solution C) twice. (2) Add solution B to solution A. (4) Add liquid B to liquid C gradually. 18°C, 130 rpm, shielded from light, mixed for 24 hours, and then allowed to acclimate with more water for 48 hours. Four changes of supplementary water were made throughout this time. (5) Adjust the coupling agent to the proper concentration using more water, then use an ozone spectrophotometer to examine it.
In vitro, NSP-SA-NHS is used as a chemiluminescent labeling reagent for immunoassays and other detection systems. The compound is conjugated to antibodies, antigens, or other biomolecules via the NHS ester group, which reacts with primary amines to form stable amide bonds. The labeled biomolecules are then used in chemiluminescent immunoassays. In the presence of hydrogen peroxide and alkaline conditions, the acridinium ester undergoes oxidation, producing light at 430 nm. The intensity of the light emission is proportional to the amount of labeled biomolecule, which is in turn proportional to the concentration of the target analyte. A rapid and sensitive chemiluminescent immunoassay of Bisphenol A with NSP-SA-NHS-labeled has been developed. |
| ln Vivo |
In vivo studies of NSP-SA-NHS are not applicable, as the compound is used as an in vitro research reagent for immunoassays and is not intended for in vivo use. The compound is not administered to animals or humans. It is used exclusively in biochemical and diagnostic assays. No extensive in vivo pharmacological or toxicological studies have been reported.
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| Enzyme Assay |
For chemiluminescent immunoassay development, NSP-SA-NHS is conjugated to antibodies, antigens, or other biomolecules. The compound is dissolved in an appropriate solvent (e.g., DMSO or DMF) and reacted with the biomolecule in a buffer at pH 7.0-8.5 for 1-2 hours at room temperature. The NHS ester reacts with primary amines on the biomolecule to form stable amide bonds. The labeled biomolecule is purified by dialysis, size exclusion chromatography, or other methods to remove unreacted compound. The labeled product is characterized by measuring the incorporation ratio (label-to-protein ratio) and chemiluminescent activity. The labeled biomolecule is then used in immunoassays, where it binds to the target analyte and generates a chemiluminescent signal upon addition of trigger solution (H₂O₂ and NaOH).
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| Cell Assay |
For cellular studies, NSP-SA-NHS is not typically used in cell-based assays, as it is a labeling reagent for immunoassays. However, it may be used to label cell surface proteins or other cellular components for detection purposes. Cells are cultured and incubated with NSP-SA-NHS-labeled antibodies or other probes. The labeled cells are then analyzed for chemiluminescent signal. No specific cellular protocols are reported.
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| Animal Protocol |
Standard in vivo protocols for NSP-SA-NHS are not applicable, as the compound is used as an in vitro research reagent. It is not administered to animals. No specific in vivo protocols are reported.
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| ADME/Pharmacokinetics |
Pharmacokinetic data for NSP-SA-NHS are not applicable, as the compound is used as an in vitro research reagent and is not intended for in vivo use. The compound is not administered systemically, and no ADME studies have been reported.
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| Toxicity/Toxicokinetics |
Toxicological data for NSP-SA-NHS are limited, as the compound is used as a research reagent. The compound is generally considered to have low toxicity at the concentrations used for labeling and immunoassays. No acute toxicity, mutagenicity, or carcinogenicity data have been reported. As with all chemical reagents, appropriate safety precautions should be taken when handling NSP-SA-NHS, including the use of personal protective equipment and work in a well-ventilated area.
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| References | |
| Additional Infomation |
NSP-SA-NHS (CAS 199293-83-9) is an acridinium ester used for chemiluminescent immunoassay. It has a molecular formula of C₃₂H₃₁N₃O₁₀S₂ and a molecular weight of 681.73. The compound is a highly sensitive chemiluminescent labeling reagent for antigens, antibodies, proteins, and nucleic acids. It emits light at 430 nm when oxidized in alkaline medium. Its stability, activity, and sensitivity exceed those of certain radioisotopes. A rapid and sensitive immunoassay for Bisphenol A has been developed using NSP-SA-NHS. The compound is strictly for research use only.
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| Molecular Formula |
C32H31N3O10S2
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| Molecular Weight |
681.733
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| Exact Mass |
681.145
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| CAS # |
199293-83-9
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| PubChem CID |
59642932
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| Appearance |
Light yellow to yellow solid powder
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| LogP |
4.842
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
10
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| Rotatable Bond Count |
12
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| Heavy Atom Count |
47
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| Complexity |
1330
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CC1=CC=C(C=C1)S(=O)(=O)N(CCCC(=O)ON2C(=O)CCC2=O)C(=O)C3=C4C=CC=CC4=[N+](C5=CC=CC=C53)CCCS(=O)(=O)[O-]
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| InChi Key |
KQFCNGKUXYNDPF-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C32H31N3O10S2/c1-22-13-15-23(16-14-22)47(43,44)34(20-6-12-30(38)45-35-28(36)17-18-29(35)37)32(39)31-24-8-2-4-10-26(24)33(19-7-21-46(40,41)42)27-11-5-3-9-25(27)31/h2-5,8-11,13-16H,6-7,12,17-21H2,1H3
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| Chemical Name |
3-[9-[[4-(2,5-dioxopyrrolidin-1-yl)oxy-4-oxobutyl]-(4-methylphenyl)sulfonylcarbamoyl]acridin-10-ium-10-yl]propane-1-sulfonate
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
DMSO : ~11.36 mg/mL (~16.66 mM)
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.4669 mL | 7.3343 mL | 14.6686 mL | |
| 5 mM | 0.2934 mL | 1.4669 mL | 2.9337 mL | |
| 10 mM | 0.1467 mL | 0.7334 mL | 1.4669 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution 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 stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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