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Created with Fabric.js 1.4.5 Analysis of different animal poisons and obtention of toxic peptides. Crotalus Durissus (Rattlesnake),Latrodectus Mactans (American Black Widow),Centruroides Limpidus (Morelos Scorpion),Rhopalurus Junceus (Blue Scorpion),Centruroides Tecomanus (Colima Scorpion)and Centruroides Suffusus (Durango Scorpion). Evaluated venoms HPCL-RPC.Electrophoresis of proteins in polyacrylamide gels. Methods for obtaining peptides Peptide characterization methods Dragendorff assay.Meyer test.Shinoda tes.Ninhydrin assay.Carbohydrates test.Sugars test. Lipids assay.Ferric Chloride test. Analysis of toxic peptides effects from different species over MCF-7 tumor human cell line. On this test was analyzed the effect of each one of the peptides from venoms obtained over the tumor human cell line MCF-7 (ATCC Number HTB-22); established according to the protocol for this cell line, Observing with a microscope and equipment Organon Teknica No.510 the cancer cell growth inhibition. Test description Analysis of inhibition capacity of TETEBENE over tumor cell lines. MCF-7. T47D.HeLa.MDA-MB-175-VII.4T1.HCC70. Evaluated tumor cell lines Solubilization and dissolution of the sample.Cell inoculation.Addition and incubation of the extracts of the samples.Test B Sulforhodamine.Determination of apoptosis by flow cytometry methods. Stages Calculation of final concentration Ctf = (Ratio1/Ratio0) x C0.Where:Ratio1 = Nt / NRefBBtRatio0 = N0 / NRefB0Ctf = Concentration in cells / ml after a time t. Final concentration. Nt = Number of events analyzed cell population after a time t. NRefBt = Number of events analyzed microparticles after time t. N0 = Events initial analysis of the cell population over time t. NRefB0 = Events analysis of the microparticles during time t.C0 = Initial concentration in cells / ml.Ctf = (Nt/N0) x C0. Simplified formula Calculation of Apoptotic index The apoptotic index allows determining the proportion of apoptotic cells which remain in detectable levels of expression for CD146, CD133 and CD117 and remain within the crop; this represents approximately 450 molecules / cell for flow FaCScan BD FACSVerse.AI=A+ / (A+ + A-)Where:AI = Rate of apoptosisA+= Cells displaying apoptotic state, represented in percentage. (annexinV binding +)A- = Cells lacking apoptotic state represented in percentage. (anexinnV-) Apoptosis rate allows obtaining the ratio of cells relative to the initial population of cells seeded have suffered apoptosis. In this way we can measure the total number of viable cells, the number of cells originally cultivated and maintained the expression of CD146, CD133 and CD117 antigens; and those who have lost this expression. Through these calculations can be obtained the total number of cells that underwent fragmentation as apoptotic bodies.CF = CS CCWhere:CF = Cells where nuclear fragmentation occurs in apoptotic bodies.CS = Initially seeded cells.CC = Cells remaining in the culture.AR = (CS CV) / CSWhere:CV = Number of viable cells. Calculation of Apoptotic rate Other calculations Growth rate.Increase in cell death.Survival rate.Nonparametric tests Mann-Whitney U.Wilcoxon rank sum test.IC50 (Calculation of IC50 was done with the dose-response curve obtained through the Organon Teknika No.510 for each cell line using the linear regression method with change in both axes) Design of molecular nanotransporters in the application of TETEBENE. Western Bloat for PARP during the aplication of TETEBENE. PARP-1 and PARP-2 proteins were placed at 1: 200dilution in 12 ml of albumin solution in 40 ml TBS for one hour under stirring. Was placed in 2ml membrane luminescent substrate to start the developing step, which was carried out in dark room by placing the membrane in revealing plate covered with plastic and exposed to Kodak film for one minute. The amount of protein was quantified by measuring the bands and densitometric analysis.
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