| Size | Price | Stock | Qty |
|---|---|---|---|
| 100mg |
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| 500mg |
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| Other Sizes |
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| ln Vitro |
The amid link between MTX and BSA as well as the amidic bond within the BSA structure can be broken by the enzyme proteinase K[2]. Guidelines (This is our suggested protocol; it should be adjusted to meet your unique needs; it is merely meant to serve as a guide). Without the need for RNA extraction, the SARS-COV2 viral component can be directly detected from inoculation viral transport medium (VTM) using proteinase K, which has better protein-degrading capabilities[3]. The steps involved in the rRTPCR detection protocol for SARS-CoV2 are as follows: 1. Divide the VTM of each SARS-COV2 sample into three 45 μL EP tubes; 2. Add 2.5 µL (20 mg/mL) concentrated proteinase K (PK), resulting in a final concentration of PK ~1 mg/ml VTM; 3. Incubate at 25 °C for 5 min; 4. Incubate at 60 °C for 15 min; 5. Incubate at 98 °C for 10 min, and store the samples at 4 °C. 6. Use 5 µL of extracted RNA as template to carry out the rRTPCR assay; 7. Utilize the BIORAD CFX96™ Real-Time Thermocycler for rRTPCR detection, with a positive standard of threshold cycle (Ct) values less than 40. Proteinase K raises the concentration of nucleic acids recovered from sputum samples and enhances their homogenization effects and positive rates[4]. The procedure for extracting influenza A virus (IAV) nucleic acid is as follows: 1. Divide the specimen into three 500 μL EP tubes; 2. Add 500 μL of Proteinase K buffer (0.4 mg/mL); 3. Incubate at 55 °C for 25 minutes, vortexing every five minutes; 4. Extract RNA using the Automated Nucleic Acid Extraction System (Zhijiang Biotechnology Co., Ltd., Shanghai, China) using 200 μL samples; 5. Assess the A260/A280 ratio of pure RNA products, using a standard range of 1.8 to 2.0.
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| References |
[1]. W Ebeling, et al. Proteinase K From Tritirachium Album Limber. Eur J Biochem . 1974 Aug 15;47(1):91-7.
[2]. Nosrati H, et al. Multifunctional nanoparticles from albumin for stimuli-responsive efficient dual drug delivery. Bioorg Chem. 2019 Jul;88:102959. [3]. Shukla A, et al. Vitality of Proteinase K in rRTPCR Detection of SARS-CoV2 Bypassing RNA Extraction. Front Cell Infect Microbiol. 2021 Nov 3;11:717068. [4]. Yu F, et al. Comparative Evaluation of Three Preprocessing Methods for Extraction and Detection of Influenza A Virus Nucleic Acids from Sputum. Front Med (Lausanne). 2018 Mar 2;5:56. |
| Molecular Formula |
C29H27N2O12P
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|---|---|
| Molecular Weight |
626.5046
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| CAS # |
39450-01-6
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| SMILES |
P(=O)(O[H])(O[H])O[C@@]12[C@@](C([H])([H])[H])(C(=C(C(=O)OC([H])([H])C3C([H])=C([H])C(=C([H])C=3[H])[N+](=O)[O-])N1C([C@@]2([H])[C@@]([H])(C([H])([H])[H])O[H])=O)OC1C([H])=C([H])C([H])=C([H])C=1[H])OC1C([H])=C([H])C([H])=C([H])C=1[H]
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| 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) |
H2O: 25 mg/mL
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: 50 mg/mL (Infinity mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.
(Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.5962 mL | 7.9808 mL | 15.9617 mL | |
| 5 mM | 0.3192 mL | 1.5962 mL | 3.1923 mL | |
| 10 mM | 0.1596 mL | 0.7981 mL | 1.5962 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.
Products are for research use only; We do not sell to patients
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