Reagents and materials:
Complete medium (CCM): α-MEM: α-low-limit basal medium containing glutamine, no nucleotide or deoxynucleotide; addition: 20% additional L-glutamine 2mmol/L, FBS The hybridoma was purified, non-heat inactivated, penicillin 100 U/ml, and streptomycin 100 μg/ml. Filtration sterilization. Store at 4 ° C for no more than 2 weeks;
FDM (fat differentiation medium): 1 FDM: CCM contained 0.5 μmol/L IBMX, 50 μmol/ml IM and 0.5 μmol/L dexamethasone. 2 Isobutylmethylxanthine (IBMX), 1000× stock solution, made up to 0.5 mmol/L with methanol. 3 Indomethacin (IM): 1000× stock solution, made up to 50 mmol/L with methanol. 4 dexamethasone: 1000 × stock solution, formulated with water to make 0.5mmol / L;
PBSA;
Trypsin/EDTA;
Polypropylene centrifuge tube, 15ml and 50ml;
Tissue culture dish, 6 wells, with a pore area of ​​9.6 cm 2 ;
0.85% trypan blue salt solution;
Neutral formalin buffer, 10%;
ORO working solution: ORO is made up of 1% and 3 parts with isopropyl alcohol. PBSA, 2 servings. 70 μm cell strainer filtration;
Improved Neubauer blood cell counter;
experimental method:
Collecting MSCs from monolayer cells;
2 ml of CCM was added to each well of a 6-well culture plate for a total of 1 x 105 cells (for human or rat). The final density is about 1 × 103 cells / cm 2;
Mark the top 3 holes as "fat" and the bottom 3 holes as "negative";
Incubate at 37 ° C, 5% CO 2 , and change the medium every two days until the cells reach 7.%-80% confluence;
When the desired cell density is reached, a complete prototype is aspirated from the well, 2 ml of FDM is added to the wells of the 6-well culture plate, and 2 ml of CCM is added to the lower row of wells;
Incubate at 37 ° C, 5% CO 2 and replace the medium every two days. Cells were stained on day 21 for ORO detection;
The differentiated MSCs were taken out from the incubator, and the monolayer cells in each well were washed twice with 2 ml of PBSA;
2 ml of formalin was added to each well, and monolayer cells were fixed at room temperature for 10 min;
The formalin was aspirated, and 2 ml of PBSA was added to each well for two times, and then washed with 2 ml of distilled water;
Add 2ml ORO working solution and incubate for 30min at room temperature;
Excess PBSA rinses each well until background staining in the "negative" well is maximally cleared;
The degree of staining of the lipid droplets was evaluated by observing the monolayer cells labeled "adipogenic" with a microscope. When the adipogenic differentiation reaches a satisfactory level, the monolayer cell ORO staining area exceeds 30%. At this point, monolayer cells can be stored for up to 7 days in 4 ° C PBSA. Alternatively, the ORO is extracted from the stained monolayer cells and assayed using the protocol previously described;
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You are always welcome to visit our company and taste our products. Contact: MS. Sunny Wang |
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| Product Description | |
| Name | Canned Tuna |
| Flavor | Brine, Oil, Salad, Chili |
| Type | Shred, Flake, Chunk, Solid |
| Certificates | EU, FDA, BRC, HALAL,HACCP,KOSHER |
| Net weight | 140g, 160g, 170g, 185g, 200g, 1kg, 1.88k. |
| Brand | Our brand or OEM, ODM |
| Shelf life | 3/4 Years |
| MOQ | 1X20'FCL |
| Payment terms | T/T, L/C |
| Delivery time | 25 days after label artwork confirmed and advance payment done. |
| Packing | normal lid or easy open,paper label or lithio can, paper carton or shrinked by tray |
| EU NO. | 3302/01034 |
| RUSSIA NO. | 3302/01034 |
| Shipping docs | Commercial Invoice |
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Packing List |
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Bill of Lading |
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Certificate Of Origin/ Form A |
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Health Certificate |
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Veterinary Certificate |
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Catching certificate |
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Or as per customer`s request |
Tuna In Pouch,Pouch Tuna Chunk,Canned Tuna Fish,Pouch Pack Tuna
Tropical Food Manufacturing (Ningbo) Co., Ltd. , https://www.tropical-food.com