MTS Cell Proliferation and Cytotoxicity Detection Kit

Chinese nameMTS Cell Proliferation and Cytotoxicity Detection Kit

English name: MTS Cell Proliferation and Cytotoxity Detection Kit

Product specifications: 500 times
Product Code: BJ-01X6321

MTS is a novel methyl sulfide compound, which belongs to the same family of tetrazolium derivatives as MTT. MTS can be degraded by mitochondrial dehydrogenase in living cells to produce brownish yellow water-soluble formazan. By measuring the spectral absorption of formazan, the proliferation of cells can be determined.

Cell culture operating procedures:
1、 Preparation of culture medium and culture cryopreservation conditions:
1) Prepare F-12K culture medium; High quality fetal bovine serum, 10%; Double antibody, 1%.
2) Cultivation conditions: Gas phase: air, 95%; Carbon dioxide, 5%. Temperature: 37 ℃, humidity in the incubator is 70% -80%.
3) Cryovial solution: 90% serum, 10% DMSO, ready to use and prepare.
2、 Cell processing:
1) Resuscitate cells: Quickly place a cryovial containing 1mL of cell suspension into a 37 ℃ water bath (with the water level lower than the lid of the cryovial) and shake to thaw. Transfer the cryovial into a 15ml centrifuge tube containing 4mL of culture medium prepared in advance and mix well. Centrifuge at 1000RPM for 4 minutes, discard the supernatant, add 1mL of culture medium and blow evenly. Then transfer all cell suspensions into a culture bottle containing 5ml of culture medium and culture overnight. The next day, change the solution and check the cell density.
2) Cell passage: If the cell density reaches 80% -90%, passage culture can be carried out.

Cell culture method:
1. Cell passaging: When the cell density reaches 80-90%, it can be passaged
① Discard the culture supernatant and wash 1-2 times with PBS or physiological saline;
② Add 2ml of 0.25% (T25 bottle) to cover the entire bottle or dish, cover it, and place it in the incubator for digestion;
③ After 1-2 minutes, observe the cells under a microscope. If most of the cells retract and a small amount of cells fall off, gently blow them to confirm digestion and add * culture medium to terminate digestion; If the cells are still attached to the wall, put them back in the incubator and continue digestion until they can be gently blown off;
④ Centrifuge the cell suspension at around 1000 RPM for 4 minutes and discard the supernatant;
⑤ Resuspend with fresh culture medium and add it to the culture bottle or dish. Add 6-8ml of culture medium to the T25 culture bottle;
⑥ The suspended cells were collected directly by centrifugation, and the cell pellet was resuspended before being transferred to new culture bottles.
2. Cell revival:
① Quickly shake and melt the cryovial in 37 ℃ warm water for about 1 minute, then add 4-5ml of culture medium and mix well.
② Centrifuge at around 1000RPM for 4 minutes, discard the supernatant, add 1-2ml of culture medium and blow evenly. Add the cell suspension to the culture bottle and add an appropriate amount of culture medium.
3. Cell cryopreservation: Perform cell cryopreservation and seed preservation when the cells are in good growth condition
① Discard the culture supernatant, wash 1-2 times with PBS or physiological saline, and add 1mL of 0.25% (T25 bottle)
② After 1-2 minutes, observe the cells under a microscope. Most of the cells retract and a small amount of cells fall off. Gently blow and pat to confirm digestion, then add * culture medium to terminate digestion;
③ Centrifuge the cell suspension at around 1000RPM for 4 minutes, discard the supernatant, and resuspend the cells in 1ml of frozen solution;
④ Place the cryovial into the program cooling box and store it in a -80 ℃ freezer. After 4 hours, transfer the cryovial to a liquid nitrogen tank for storage.

TM3 (mouse testicular interstitial cells) 5 × 106 cells/bottle × 2

TSCCa (human tongue squamous cell carcinoma cells) 5 × 106 cells/bottle × 2

U-2 OS (human osteosarcoma cells) 5 × 106 cells/bottle × 2

U251 (human glioma cells) 5 × 106 cells/bottle × 2

U-87 MG (human glioma cells) 5 × 106 cells/bottle × 2

U-937 (human histiocytic lymphoma cells) 5 × 106 cells/bottle × 2

V79 (hamster lung cells) 5 × 106 cells/bottle × 2

VCaP (human prostate cancer cells) 5 × 106 cells/bottle × 2

Vero (African green monkey kidney cells) 5 × 106 cells/bottle × 2

WI-38 (human embryonic lung cells) 5 × 106 cells/bottle × 2

WISH (human amniotic membrane cells) 5 × 106 cells/bottle × 2

Y1 (Y-1) (mouse adrenal cortex cells) 5 × 106 cells/bottle × 2

YAC-1 (mouse lymphoma cells) 5 × 106 cells/bottle × 2

ZR-75-1 (human breast cancer cell) 5 × 106 cells/bottle × 2

ZR-75-30 (human breast cancer cell) 5 × 106 cells/bottle × 2

16HBE (human bronchial epithelial like cells) 5 × 106 cells/bottle × 2

801-D (human giant cell lung cancer cells) 5 × 106 cells/bottle × 2

A2 (human adenoid cystic carcinoma cells) 5 × 106 cells/bottle × 2

A-427 (human lung cancer cells) 5 × 106 cells/bottle × 2

A-498 (human kidney cancer cells) 5 × 106 cells/bottle × 2

A875 (human melanoma cells) 5 × 106 cells/bottle × 2

Lung alpha/beta hydrolase protein 3 antibody

Adenosine triphosphate binding cassette transporter F subunit 3 antibody

Adenosine triphosphate binding cassette transporter A subunit 5 antibody

ADCK1 protein antibody

Acyl binding domain protein 4 antibody

Alpha/β hydrolase 4 antibody

ADCK2 protein antibody

Aldehyde dehydrogenase 16 family A1 antibody

Adhesive molecule IgG like domain protein 3 antibody

Ethanol dehydrogenase 1 antibody

Delta aminolevulinic acid dehydratase antibody

Sarcopenia associated protein 4 antibody

Alpha 2 macroglobulin antibody

ACCSL protein antibody

Alpha fetoprotein antibody

β - amyloid precursor protein binding protein 3 antibody

Star shaped actin antibody

Systemic lupus erythematosus associated protein TREX1 antibody

Phosphorylated systemic lupus erythematosus prequelin TREX1 antibody

Phosphorylated systemic lupus erythematosus associated protein TREX1 antibody

MTS Cell Proliferation and Cytotoxicity Detection KitHead associated protein complex AP-2 μ chain 1 antibody

Phosphorylated linker associated protein complex AP-2 μ chain 1 antibody

Membrane adhesion protein 7 antibody

Operating Procedures
1. Add 100 μ L of cells per well to a 96 well plate. Typically, for cell proliferation experiments, add 100 μ L of 2000 cells per well. For cytotoxicity experiments, add 100 μ L of 5000-10000 cells per well (the specific number of cells used per well depends on cell size, proliferation rate, etc.). Cultivate cells in a 37 ℃ 5% CO2 incubator for 24 hours
2. Add 0-10 μ L of the test compound at an appropriate concentration.
3. Incubate the 96 well plate in a cell culture incubator containing 5% CO2 air and 100% humidity at 37 ℃ for an appropriate period of time.
4. Dilute 5 x MTT with diluent to 1 x MTT solution.
5. Add 50 μ L of 1 × MTT solution to each well and incubate at 37 ℃ for 4 hours to reduce MTT to formamide.
6. Suck out the supernatant, add 150 μ L DMSO to each well to dissolve formaldehyde, and shake well with a shaker.
7. The enzyme-linked immunosorbent assay (ELISA) detects the optical density of each well at a wavelength of 570nm (if there is no 570nm filter, a 560-600nm filter can be used).
8. Result analysis
a、 Cell survival rate: Subtract the background OD value (* culture medium plus MTT, no cells) or blank drug well OD value (* culture medium plus different dilutions of the test drug plus MTT, no cells) from the OD values of each test well, and take the average OD value of each replicate well ± SD. The cell survival rate is expressed as T/C%, where T is the OD value of the drug added cells and C is the OD value of the control cells. Cell survival rate%=(OD of drug treated cells/OD of control cells) × 100
b、 Calculate the drug concentration (IC50) at T/C=50% and the drug concentration (IC90) at T/C=10%