Size | Price | Stock | Qty |
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100mg |
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250mg |
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500mg |
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1g |
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Other Sizes |
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ln Vitro |
Collagenase, Type III is used for the preparation of primary breast cells.
Preparation of stock solution 1. Add 1 mL of HBSS (Hank equilibrium salt solution, containing Ca2+and Mg2+) to 100 mg of collagenase, shake to fully dissolve, and prepare a storage solution of 100 mg/mL (i.e. 100 x); 2. Use low protein binding 0.22 μ M filter membrane filtration for sterilization, immediately use or divide into small portions, and freeze in the dark at -20 ° C to -5 ° C; 3. Thaw on ice before use to avoid repeated freeze-thaw cycles. The commonly used concentration for tissue and cell dispersion is 0.5-2.5 mg/mL, and the commonly used concentration for cartilage digestion is 1-2mg/mL. The optimal working concentration required needs to be determined based on specific experimental conditions or reference to relevant literature. Separation of Organ/Tissues 1. Use sterile surgical knives or scissors to cut tissue into 3-4 mm sized tissue blocks; 2. Wash tissue blocks several times using HBSS containing Ca2+and Mg2+; 3. Add sufficient HBSS containing Ca2+and Mg2+to immerse the tissue block, and add collagenase to the required working concentration; 4. Incubate at 37 ° C for 4-18 hours. Using a horizontal shaker and supplementing digestion with 3 mM CaCl2 during digestion can improve digestion efficiency; 5. The dispersed cells can be screened using stainless steel or nylon mesh, and collected for later use. Add an appropriate amount of fresh collagenase working solution to the partially dissociated tissue and continue incubation at 37 ° C for further digestion; 6. Wash the collected cells several times using HBSS without collagenase; 7. Resuspend the above cells in cell culture medium and calculate the density of live cells using an automatic cell counter or other methods; 8. Inoculate cells using appropriate cell culture medium on a cell culture dish. Organ perfusion 1. Adding collagenase to HBSS containing Ca2+and Mg2+preheated at 37 ° C, and adding 3 mM of CaCl2 can help improve separation efficiency; 2. Inject collagenase working solution into the corresponding organs at the optimized rate; 3. The recovered infusion solution from the above process is passed through a stainless steel or nylon mesh screen to separate the dissociated cells or small fragments of tissue from larger clumps. The tissue that has not been fully decomposed needs to be further incubated with fresh collagenase working solution at 37 ° C. 4. Same as steps 6-8 in the above section "Separation of Organ/Tissues". |
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References |
Exact Mass |
812.418
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CAS # |
9001-12-1
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Appearance |
Yellow to brown solid
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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 |
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: ≥ 100 mg/mL
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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.) |
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.