Boc-Val-Leu-Gly-Arg-pNA |
Katalog-Nr.GA21162
One-Click Copy Product Info
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Boc-Val-Leu-Gly-Arg-pNA is a chromogenic peptide substrate. Boc-Val-Leu-Gly-Arg-pNA can be specifically hydrolyzed by the limulus clotting enzyme, releasing p-nitroaniline (pNA) to produce a color change.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 68223-95-0
Sample solution is provided at 25 µL, 10mM.
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Boc-Val-Leu-Gly-Arg-pNA is a chromogenic peptide substrate. Boc-Val-Leu-Gly-Arg-pNA can be specifically hydrolyzed by the limulus clotting enzyme, releasing p-nitroaniline (pNA) to produce a color change. Boc-Val-Leu-Gly-Arg-pNA can be used for quantitative detection of endotoxins and in research related to limulus amebocyte lysate (LAL) reagents[1-3].
References:
[1] Bahgat M, Aboul-Enein MN, El Azzouny AA, et al. A cyclohexanecarboxamide derivative with inhibitory effects on Schistosoma mansoni cercarial serine protease and penetration of mice skin by the parasite. Acta Pol Pharm. 2009 May-Jun;66(3):333-40.
[2] Bahgat M, Khalifa N. Differential inhibitory effect of newly synthesized pyridine-2-one derivatives on the cercarial serine protease activity of the parasite Schistosoma mansoni. Acta Pol Pharm. 2006 May-Jun;63(3):181-8.
[3] Iwanaga S, Morita T, Harada T, et al. Chromogenic substrates for horseshoe crab clotting enzyme. Its application for the assay of bacterial endotoxins. Haemostasis. 1978;7(2-3):183-8.
Method for Determining Bacterial Endotoxins Using Boc-Val-Leu-Gly-Arg-pNA[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 the synthetic substrate Boc-Val-Leu-Gly-Arg-pNA in pyrogen-free distilled water and dilute to the working concentration (0.1mM or 0.5mM). Prepare 0.1M Tris-HCl buffer (pH=8.0). Prepare a 0.5M MgCl₂ solution. Dissolve standard endotoxin (E. coli 0111-B4 lipopolysaccharide) in pyrogen-free normal saline to prepare a solution of the desired concentration (0.1%). Prepare limulus (Tachypleus or Limulus) amebocyte lysate.
2. The sample to be tested (e.g., bacterial endotoxin solution) must be prepared and diluted using sterile, pyrogen-free glassware and reagents.
3. Endotoxin Detection: Sequentially add 0.8ml of the substrate working solution dissolved in Tris-HCl buffer (pH=8.0), 50µl of 0.5M MgCl₂ solution, and 20µl of the endotoxin sample solution to the reaction system. Pre-incubate the mixture at 37°C for 3 minutes. Subsequently, add 20-50µl of the limulus amebocyte lysate to initiate the reaction and continue incubation at 37°C for 15 minutes (or extend as needed to 30, 60, or 240 minutes). After incubation, add 100µl of glacial acetic acid to stop the reaction. Use a spectrophotometer to measure the absorbance of the released p-nitroaniline (pNA) at 405nm wavelength. The absorbance value is proportional to the endotoxin concentration.
Precautions:
(1) To ensure accurate results, the entire experimental process must use sterile, pyrogen-free utensils and solutions. Some substrates may require organic solvents for dissolution; please pay attention to operational safety.
(2) For your safety and health, please wear a lab coat and disposable gloves during operation.
References:
[1] Iwanaga S, Morita T, Harada T, et al. Chromogenic substrates for horseshoe crab clotting enzyme. Its application for the assay of bacterial endotoxins. Haemostasis. 1978;7(2-3):183-8.
| Cas No. | 68223-95-0 | SDF | |
| Formula | C30H49N9O8 | M.Wt | 663.78 |
| Löslichkeit | Soluble in DMSO | 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 | |||
|
1 mg | 5 mg | 10 mg |
| 1 mM | 1.5065 mL | 7.5326 mL | 15.0652 mL |
| 5 mM | 301.3 μL | 1.5065 mL | 3.013 mL |
| 10 mM | 150.7 μL | 753.3 μL | 1.5065 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: >98.00% Appearance: A solid
- COA (Certificate of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 31 reference(s) in Google Scholar.)