Human bcl2 elisa kit

To evaluate the inhibitory activity of the CM derivative against topo I and topo II enzymes, human DNA topoisomerase 1 ELISA kit (#MBS289119, MyBioSource, San Diego, CA, USA) and human DNA topoisomerase II ELISA kit (#EKN48879-96T, BIOMATIK, Wilmington, DE, USA) were utilized. Also, to evaluate the level of bax (proapoptotic factor) and bcl2 (antiapoptotic factor) proteins, (#E-EL-H0562, human bax ELISA kit, Elabscience, TX, USA) and (#E-EL-H0114, human bcl2 ELISA kit, Elabscience, Houston, TX, USA) were utilized, respectively. Cancer cells were treated with the IC50 of the CM derivative for 48 h. Then, untreated and treated cells were harvested followed by measurement of topo I, topo II, bax, and bcl2 according to manufacturer’s instructions. A standard curve was obtained, and the absorbance was measured at 450 nm. The concentration of topo I, topo II, bax, and bcl2 were calculated from the standard curve. The experiment was performed in triplicate.

The concentration of protein in the cultured cells was measured using an ELISA human immunoassay kit: Human Bcl-2 ELISA Kit (Elabscience, Wuhan, China, catalog no. E-EL-H0114) and Quantikine ELISA Human BDNF Immunoassay (R&D Systems, McKinley Place, MN, USA catalog no. DBD00). The absorbance was read at 450 nm using an ELX 800 IU automated Microplate Reader (Bio-Tek Instruments, Inc., Winooski, VT, USA; Gene 5 Software V 3.02). The results were analyzed using a quadratic log–log curve fit.