Electrophoresis of Proteins from Cauliflower Fractions by SDS-PAGE Laboratory Summary Electrophoresis is a technique where molecules are separated according to their physical properties such as size‚ charge‚ and/or shape. Charged proteins are commonly separated in this matter using PAGE (polyacrylamide gel electrophoresis) to identify individual proteins present in samples. In this lab‚ SDS-PAGE was used. SDS-PAGE separates charged proteins primarily by size because the ionic detergent sodium dodecyl
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experiment‚ an absolute standard was calculated using BSA‚ so that the concentrations of the other unknown protein solutions can be determined (Lambert et.al‚ 2011).The different assays used for this protein quantification were Lowry‚ Bradford (Coomassie Blue) and UV direct. Protein assays help to determine the amount of desired particle
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Objectives Protein Isolation: Protein isolation for a western blot uses detergents and mechanical force to separate seeded cells from their container. Eukaryotic cells are attached to the surface of a flask by cadherins. In the past‚ we’ve separated the cells from the flask by breaking these bonds with a protease‚ but in order to keep the proteins intact‚ a different method needs to be used to extract the proteins. In protein extraction for a western blot‚ we use detergents to lyse the membrane
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amounts of Bovine Serum Albumin (BSA). Varying amounts of BSA at a concentration of .5µ/µl were mixed with the water and 1mL of coomassie dye. The equation y=0.0259x +0.0511 represents the best fit line for a data set taken from µg of protein equals two to ten µg. Optical density (OD) was surveyed using a Mach V visual spectrophotometer at 595. Worktable for Coomassie Assay Tube # | Sample ID | Water (µl) | Sample (µl) | Protein (µg) | OD @ 595 | 1 | BLANK | 1000 | 0 | 0 | 0 | 2 | STAN
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Calculations are as followed: Sample (Tube) A was prepared with pure dH2O since the concentration of protein was 0 μg/mL. Upon addition of Coomassie Blue G-250 and the completion of the incubation period it was noticeable that solutions of higher concentration had a deeper color blue opposed to those with a lower concentration. The absorbance range for the standard curve (figure 1) was determined
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Background: Lipid and protein composition varies in tissue types. Results: When comparing liver‚ muscle‚ and adipose tissue‚ adipose tissue had the highest percent lipid and liver tissue had the highest percent protein. Conclusion: Based on analysis of the gels‚ the liver appeared to have the most variety in proteins when comparing liver‚ muscle‚ and adipose tissue. Significance: In order to determine the composition of a tissue‚ specific macromolecules can be extracted‚ quantified‚ and analyzed
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In this lab we employed various assays utilizing a biuret reagent‚ coomassie brilliant blue reagent‚ and ultraviolet light in order to determine the identity of six unknown solutions and the concentration of a bovine serum albumin sample. We were given three samples that lacked protein‚ and three samples containing proteins‚ and using a spectrophotometer we assessed the amount of light absorbed versus the light transmitted‚ based on the principles of the Beer-Lambert Law. The three proteins used
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Beta-Galactosidase and Western Blot 0 1. EMG 9 and EMG 26 contain strain _lac-_(I- Z+ Y+) and strain _lac -_ (I+ Z- Y-)respectively.Three genes huddled together on the chromosome are required for two strains of _E.coli_ to utilize lactose.Consisting of three genes‚ namely‚ _lacZ_‚ _lacY_ and _lacA_‚ the _lac_ operon orderly handles these genes to code specific enzymes necessary for the metabolism of lactose. The genes _lacZ_‚ _lacY_ and _lacI_ would code for beta-galactosidase‚ galactosidase permease
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The Bradford protein assay is a colorimetic assay based on the interaction of the blue dye “Coomassie” and the protein. The blue ye firsts donates free electron to the protein and causes the protein to lose its native state and exposing it hydrophobic pockets. These hydrophobic pockets then binds to the Coomassie dye through Van Der Wall forces. The binding protein then stabilizes the blue form of the Coomassie dye. Hence‚ the absorbance reading can be obtained and used to measure the protein concentration
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Expressing and Purifying the Recombinant form of Green Fluorescent Protein (rGFP) from the E.coli strain using Ni2+ agarose affinity chromatography technology Abstract The purpose of this experiment was to express and purify the his6-tagged recombinant form of GFP (rGFP) from the organism E.coli using Ni2+ agarose affinity chromatography. The expression of rGFP was confirmed qualitatively using the UV light and was expressed in the E.coli strain BL21 (DE3) (-- removed HTML --) (-- removed HTML
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