MATERIAL AND METHODS Recombinant protein production The recombinant T-Rp3 was expressed in E. coli BL21 (DE3) as previously described (Favaro et al.‚ 2014) and purified by a single step His-based affinity chromatography in an ÄKTA purifier FPLC (GE Healthcare). Protein was then dialyzed against 50 mM MES buffer (pH 6‚0) overnight at room temperature‚ frozen in liquid nitrogen and stored at -80 ˚C after 0.22 μm pore membrane filtration. Evaluation of DNA-protein interaction by gel retardation
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The following equations have to be applied in order to calculate the number average molecular weight ( Mn )‚ the weight average molecular weight ( Mw ) and the polydispersity index ( PDI ) : _ Mn = ΣNiMi / ΣNi _ Mw = ΣWiMi/ ΣWi = ΣNiMi2 / ΣNiMi _ _ PDI = Mw / Mn The molecular weight of polystyrene repeat unit (CH – CH2 )n is M = 104 C6H5 Using the input data of the 4 monodisperse fractions‚ we create the following table: Sample Wt/m Ni Mi NiMi
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Simple Diffusion 1. The following refer to Activity 1: Simulating Dialysis (Simple Diffusion). Which solute(s) were able to pass through the 20 MWCO membrane? According to your results‚ which solute had the highest molecular weight? ______________________________________ Which solute displayed the highest rate of diffusion through the 200 MWCO membrane? __________________________ Using the data from Chart 1‚ explain the relationship between the rate of diffusion and the size of the solute.
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able to detect and analyze proteins based on their ability to bind to a specific antibody‚ the SDS-PAGE and Western Blot was performed. SDS Polyacrylamide Gel Electrophoresis (SDS-PAGE) is a very common technique used to separate proteins based on molecular weight under the influence of an applied electrical field and then used to prepare for the Western Blot (#1 Lehninger). The support medium used is a polyacrylamide gel and‚ it also used sodium
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My graduate studies dealt with extensive and comprehensive advanced coursework and research‚ which not only helped me learn and understand the intricacies of Microbiology and Molecular Biology‚ but also provided an insight of its association with other fields like Biochemistry and Bioinformatics. I completed my graduate research work under the guidance of Dr. Shaikh‚ who is an expert in the field of Enzymology and Virology. The
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which could resist disasters such as flood and typhoons. With‚ the status of our school and our place here in Camarines Sur‚ we do not have the appropriate place and weather to conduct a similar research. With regards with the National Institute of Molecular Biology and Biotechnology (BIOTECH)‚ the institutions presented an overview of their researchers but not in a detailed way due to time constraints. Included of their researchers were the Virgin Coconut Oil‚ Organic Fertilizers‚ researches on bacteria
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antifreeze solution. 3. Using the formula molar mass = grams/moles calculate the molecular mass of antifreeze in the 10% solution. 4. Using the equation ΔT = Kf m¸calculate the molality of the 20% antifreeze solution. 5. Use the formula molality = moles solute/kg solvent to find the number of moles in 20% antifreeze solution. 6. Using the formula molar mass = grams/moles calculate the molecular mass of antifreeze in the 20% solution. 7. Average the molar masses you calculated
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incubated for 48 h and subjected to agarose gel electrophoresis. DNA ladder characteristic of apoptosis were observed in both cell lines (Fig. 5). No DNA fragmentation occurred in the untreated cells. 3.9. Caspase-3 expression in apoptotic cells The molecular effectors that activate a series of protease that leads to programmed cell death by intrinsic and extrinsic pathways are known as caspases. Caspase-3 dependent apoptotic cell death was induced by Pierisin-6 in human cancer cell lines were . The HeLa
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nucleic acids and proteins. Electrophoresis can take a mixture of macromolecules of different molecular weights‚ shapes‚ and various electrical charges to determine all the various compounds in the mixture and allowing for further purification that can aid in details of individual elements of the mixture being studied. Agarose gel electrophoresis is a very important technique used in the field of molecular science. The field of forensics utilizes agarose gel electrophoresis by allowing investigators
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the steps and procedures laid out for me. Results Experiment Results Predict Question: Predict Question 1: The molecular weight of urea is 60.07. Do you think urea will diffuse through the 20 MWCO membrane? Your answer : a. Yes‚ but very slowly. Predict Question 2: Recall that glucose is a monosaccharide‚ albumin is a protein with 607 amino acids‚ and the average molecular weight of a single amino acid is 135 g/mole. Which of the following will be able to diffuse through the 200 MWCO membrane
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