Sal and Apa were purchased from Meilun Biotechnology (Dalian, China). Fluorescein isothiocyante (FITC) was achieved from Sigma-Aldrich (St. Louis, MO, USA). The primary and second antibodies were obtained from Abcam (Cambridge, MA, USA). The RPMI-1640 medium and fetal bovine serum (FBS) were obtained from Gibco BRL (Carlsbad, CA, USA). The 0.25% trypsin-EDTA and penicillin, and streptomycin were purchased from Invitrogen Co., USA. The Cell counting kit-8 (CCK-8) and Annexin V-FITC Apoptosis Detection kit were bought from BD PharMingen (Heidelberg, Germany). All the other solvents were of chromatographic grade and obtained from Sinopharm Chemical Reagent Co., Ltd (Shanghai, China).
Cell counting kit 8 cck 8
Manufactured by BD
Sourced in United States
The Cell Counting Kit-8 (CCK-8) is a colorimetric assay for the determination of cell viability and cytotoxicity. It utilizes a water-soluble tetrazolium salt that is reduced by cellular dehydrogenases, resulting in the formation of a yellow-colored formazan dye. The amount of formazan dye generated is directly proportional to the number of living cells in the sample.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Trypsin edta, by Thermo Fisher Scientific (1 mentions)
Fitc annexin 5 apoptosis detection kit, by BD (1 mentions)
Stripping buffer, by Merck Group (1 mentions)
Thioglycollate, by Merck Group (1 mentions)
Antibodies specific for, by Cell Signaling Technology (1 mentions)
Phospho mtor, by Cell Signaling Technology (1 mentions)
Dulbecco s modified eagle medium dmem, by Thermo Fisher Scientific (1 mentions)
Phospho akt, by Cell Signaling Technology (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
P70s6k, by Cell Signaling Technology (1 mentions)
Phospho p70s6k, by Cell Signaling Technology (1 mentions)
Propidium iodide, by Merck Group (1 mentions)
Annexin 5 fluorescein isothiocyanate fitc assay kit, by BD (1 mentions)
Hydroxychloroquine hcq, by Merck Group (1 mentions)
Spectra flour plus, by Tecan (1 mentions)
P akt, by Affinity Biosciences (1 mentions)
6 protocols using cell counting kit 8 cck 8
1
Evaluating Cellular Response to Compounds
2
Apoptosis and Cell Signaling Assays
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Dulbecco’s Modified Eagle Medium (DMEM), fetal bovine serum (FBS), and penicillin/streptomycin were obtained from Gibco (Life Technologies, NY, USA). Phosphate buffered saline (PBS), protease and phosphatase inhibitor cocktails, bovine serum albumin (BSA), Radio-Immunoprecipitation Assay (RIPA) lysis buffer, stripping buffer, propidium iodide (PI), and thioglycollate were from Sigma Aldrich (St. Louis, MO, USA). An annexin V-FITC-base apoptosis detection kit, a Cell Counting Kit-8 (CCK-8), and Transwell chambers (with Matrigel pre-coating) were from BD Biosciences (San Jose, CA, USA). Antibodies specific for microtubule-associated protein 1A/1B-light chain 3 (LC3), Beclin1, P70S6K, PI3K, AKT, mTOR, phospho-AKT, phospho-mTOR, and phospho-P70S6K were from Cell Signaling (Santa Cruz, CA, USA).
3
Investigating Autophagy and Apoptosis in Cells
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The antibodies for microtubule-associated light chain 3 (LC3) (cat. no. 12741s) and cleaved-caspase-3 (cat. no. 9664s) were purchased from Cell Signaling Technology, Inc. p62 (cat. no. Ab109012), Ki67 (cat. no. Ab16667), and caspase-3 (cat. no. Ab32351) antibodies were obtained from Abcam. Anti-CD31 (cat. no. AF6191) was obtained from Affinity. Hydroxychloroquine (HCQ) was purchased from Sigma-Aldrich (Merck KGaA); Annexin V-fluorescein isothiocyanate (FITC) Assay kit was from BD Biosciences, Cell Counting Kit-8 (CCK-8, cat. no. CK04) was purchased from Dojindo Molecular Technologies, Inc., and TRIzol® (cat. no. 9109) was purchased from Thermo Fisher Scientific, Inc.
4
Cell Viability Assay Protocol
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The cell viability detection was performed according to the Cell Counting Kit-8 (CCK-8) (BD Pharmingen, USA) method [20 (link)]. Cells were seeded in 200-μL medium, and the cell density was adjusted to 5 × 103 cells per well in 96-well plate. One hundred microliters of culture medium was pipetted into the wells of 96-well plates as the blank control group. The plates were kept in an incubator at 37 °C under 5% CO2 for 24 h. Briefly, after the treatment, 10 μL of CCK-8 (Kumamoto, Japan) was added to each well and the plates were incubated for 1 h. The optical density (OD) was measured by microplate reader (Tecan; Spectra Flour Plus) at 450 nm. The procedure was carried out thrice to obtain the mean of the collected readings. The percentage of inhibition of proliferation was calculated by (1 − mean OD for drug group/mean OD for control group) × 100%.
5
Anticancer Properties of C. tinctoria
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The C. tinctoria was from the research base of the Uyghur Autonomous Region Institute of Medicinal Plants (Xinjiang, China). Human lung carcinoma (A549 and NCI-H292) cells and BEAS-2B were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). RPMI-1640 medium and fetal bovine serum (FBS) was from Gibco (United States). Penicillin/streptomycin, and dimethyl sulfoxide (DMSO) were purchased from Sigma (Missouri, United States). Cell Counting Kit-8 (CCK-8) and Annexin V-FITC/PI apoptosis detection kits were from BD Biosciences (United States). HPLC grade methanol and acetonitrile were purchased from Merck (Darmstadt, Germany). The antibodies of Caspase-3, Bax, and Bcl-2 were all obtained from Abcam (United States); those of p-PI3K, PI3K, p-Akt, and Akt were from Affinity Biosciences (Cincinnati, United States); and, that of β-actin was from Proteintech Group (Wuhan, China). An enhanced chemiluminescence (ECL) kit was purchased from PerkinElmer (Waltham, United States).
6
Exosome-induced EPC Proliferation
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EPC proliferation was evaluated using the Cell Counting Kit-8 (CCK-8) (BD Biosciences). Our team cultivated transfected cells in 96-well plates with Exos in HG conditions for 1 d in wells to which 10 μL CCK-8 reagent and 90 μL fresh culture medium was previously added. Absorbance was detected at 450 nm using a microplate reader following incubation at 37 °C for 2 h.
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