Essay Chemical Weight Of A Protein

2174 Words Nov 20th, 2016 9 Pages
In this experiment, a SDS-PAGE gel was used to analyze the protein samples from the MBP-AP and WT-AP experiments. The samples are then referenced to the ladder to determine the molecular weight of the MBP-AP and WT-AP proteins. Then the UV absorbance of the two proteins from 240 nm to 340 nm is determined using a nanovolume cuvette. The absorbance at 280 nm was then used in conjunction with data from previous experiments to determine the concentration of the MBP-AP and WT-AP protein samples. Results of experiments showed that the SDS-PAGE gel yielded expect bands and the approximate molecular weight of wild type alkaline phosphatase and maltose binding protein-alkaline phosphatase is 49 kDa and 95 kDa, respectively. Additionally, the concentrations of MBP-AP Elution 1 protein and WT-AP Stage 4 are 0.278 mg/mL and 0.582 mg/mL, respectively.

Introduction To determine the purity of a protein, a sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) is often utilized. It requires two basic components: an electrophoresis chamber, which is a gel box that contains the gel, and a power supply that provides constant and suitable current. To prepare the polyacrylamide gel, free radical polymerization of acrylamide is performed by reacting acrylamide with the cross-linking agent N,N’-methylene-bis-acrylamide. The reaction is controlled by an initiator-catalyst system, ammonium persulfate (APS)–N,N,N’,N’-tetramethylethylenediamine (TEMED). Sometimes, sodium dodecyl…

Related Documents