Bcl 2 elisa kit

Bcl-2 protein concentration was determined using the Bcl-2 ELISA kit (Invitrogen, Thermofisher Scientific, Waltham, MA, USA) according to the manufacturer’s instruction. The colon cancer cells were plated and treated for 24, 48, and 72 h at different concentrations (EC₅₀, 2XEC₅₀). The Bcl-2 concentration was according to the color intensity measured at 450 nm by using a microplate spectrophotometer (BioTek™ EON™ Microplate Spectrophotometers, Fisher Scientific, Suwanee, GA, USA). Data was presented in fold-change of absorbance from treated cells against absorbance from untreated cells as follows: $Fold change of Bcl-2= \frac{Absorbance of MS13 treated cells}{Absorbance of untreated cells \left(control\right)}$

Naringenin, isopropyl myristate, castor oil, and olive oil were purchased from Sigma-Aldrich, (St. Louis, MO, USA). Ethanol, Tween 20, Tween 80, and PEG 200 were purchased from Merck, USA. Sefsol 218 was procured from Nikko Chemicals (Tokyo, Japan), whereas Capryol 90, Transcutol HP, and Labrafac PG were obtained from Gattefosse (Saint Priest Cedex, France). The Caspase-3 Colorimetric Assay Kit (Catalog No. K106) was purchased from BioVision, USA. Human Bax ELISA (EIA-4487) was purchased from DRG International, Inc., USA. The Bcl-2 ELISA Kit (Cat. No. 99-0042) was procured from Invitrogen Corporation, CA, USA. The Thiazolyl Blue Tetrazolium Bromide (MTT) reagent kit was purchased from ABCAM, Cambridge, UK. The Annexin V-FITC Apoptosis Detection Kit and Cell cycle kits were purchased from BD Pharmingen (San Diego, CA, USA). Other chemicals used were of analytical grade.