Anti Cow Serum Lab Report

The colour of each test tube was recorded and if proteins were present that was recorded for each test tube. Finally, the pH was recorded for each sample using pH

3.8.2. Agarose Gel Electrophoresis Performed according to Brody et al. Reagents 1. Agarose 2. Tris borate EDTA (TBE) buffer, pH-8.0,1x buffer solution.

A0123942_Gel Electrophoresis Report Name: Lee Zixuan Process of Gel Electophoresis: Gel electrophoresis in this case involves the placement of both genomic and plasmid DNA inside the wells of the agarose gel, together with a gel loading buffer. The agarose gel contains mini pores such that when an electric current is switched on, it would be able to separate the bigger segments of DNA bands from the smaller ones. As DNA is negatively charged due to the phosphate group, it would move towards the positive electrode. Smaller molecules of DNA would be able to move faster than the bigger molecules of DNA. Upon completion, the separated fragments of DNA can be visualized under UV light and the distinct bands could be seen.

During the binding process the substrate requires some assistance in order to bind to the enzyme properly. This is done by several different catalytic mechanism. The most abundant catalytic mechanism is known as the general acid-base catalysis. For this reaction to occur, one of the eight amino acid residues, shown in fig 6-9, will act as a proton donor or a proton acceptor. The amino acid residues known for their acidic form will function as the proton donor and the residues that form a base will act as a proton acceptor.

Protein self-association can be triggered by chemical transformations; it is also sensitive to physical parameters such as temperature and pressure. Moreover, it is strongly affected by changes in the properties of the medium, such as, pH, the electrolyte concentration, and the presence of co solvents or additives (Stenstan et al.

• Molecular weights of protein bands in each of the three unknown samples B, C, and D were determined using the standard curve. On the Y-axis, the corresponding migration distance (cm) was found and traced to meet the line of the standard curve. This point was then traced to meet the corresponding place on the x-axis, in which molecular weight of known samples were plotted.

Antibody Detection (Antibody Screen) Three tubes were labeled 1-3 and to each patient serum was added. Group O reagent screening red cells 1 was added to tube 1, reagent screening red blood cells 2 was added to tube 2, and reagent screening red cells 3 was added to tube 3. The patient serum is the source of the antibody and the Group O reagent screening red cells are the source of the antigen in this screen.

It is commonly referred as lattice entrapment where enzyme is not bind by strong force and no structural distortion is seen. It minimizes leaching of enzymes as well as denaturation of enzymes. It also helps to create optimal microenvironment for the enzyme. Polymers, sol-gels, can be used as encapsulating agent. For example, Aluminium alignate acts as support material for Candida tropicalis in phenol

When the gel was polymerized, isopropanol was removed on the top of the running gel with water and thus, water was also removed as well. Then, stacking gel was prepared and its polymerization reaction was started with TEMED, and its solution was quickly poured on the top of the running gel. Therefore, a comb was inserted into the stacking gel before its polymerization, to form the loading wells. When the gel was polymerized, the inner chamber was filled with electrophoresis running buffer and the outer chamber was filled half. Chemical modification of ERAB (Wild Type and Mutant) Protein with Fluorescein Maleimide Wild type and mutant ERAB

5 μl of washed FVIII loaded erythrocytes diluted with 100 μl PBS incubated at 4 for 30 minutes with FVIII monoclonal antibody. Secondary antibody conjugated with FITC was added and incubated at 4 for 30 minutes again. Cells were analysed with partec

2) 5-aminolevulinic acid is transported to the cytosol for formation of porphobilinogen molecule. 3) After formation

Approximately 1µg Mbgl was used with 5 mM 4-nitropheny-β-D-glucopyranoside (PNPG) in the reaction mixture of either 2 ml or 1 ml. The reaction was stopped by adding an equal volume of 0.2M Na2CO3and the released product 4-nitrophenol was quantified based on the millimolar extinction coefficient of 18.1 mM-1cm-1at 400 nm (Workman and Day 1982). The optimum pH was determined using the same assay in the 100 mM phosphate-citrate buffer in the pH range of 3.0 to 7.0 and for pH 8.0; 100 mMtris buffer was used. Similarly, the temperature optimum was determined in 100mM citrate buffer of pH-6.0. Thermal stability was determined by incubating the protein solution at 50°C and 55°

A knowledge of the origin of the stability of proteins in aqueous solution is essential to the understanding of their structure and function. The stability of a globular protein in aqueous solution can be determined by studying the disruption of its native structure, i.e., the process of denaturation[1]. In this process the native folded protein structure is converted into a form that is predominantly unfolded but can still possess some residual folded structure[2]. Consequently, the fully unfolded or random-coil state of protein, which is the ideal reference state in discussions of the thermodynamic stability of proteins, is not always experimentally accessible[2]. Through model systems consisting of peptides or shortened proteins, it is

The MG state is a compact denatured state with a significant native like secondary structure but a largely disordered tertiary structure. In addition, there are studies demonstrating that proteins can convert from unfolded to folded or molten-globule states upon addition of large amounts of crowding agents. For instance, unfolded cytochrome c at pH 2 can adopt a molten globule structure in the presence of crowding agents, unfolded RNase A at pH 3 adopts a folded-like structure upon addition of 350 mg/ml PEG 20,000 or Ficoll 70, and the reduced and carboxyamidated form of RNase T1 that is intrinsically unstructured at pH 7 was found to exhibit some catalytical activity upon the addition of 400 mg/ml dextran 70. In addition, protein binding to a membrane surface results in “partial denaturation” (i.e. being transformed into a non-native state). The effects of various polyols, such as ethylene glycol, glycerol, erythritol, xylitol, sorbitol, and inositol, on the structure of acid-unfolded horse cytochrome c at pH 2 were investigated.

If proteins are able to be separated based on molecular size factors like intrinsic charge and molecular radious must be nutralized. Therefore, in this technique SDS (sodium dodiside sulfate) page use SDS that provides negative net charge to one another protein components at the same as they are detergent is it into linear structure. Due to these reasons factors like molecular radious and intrinsic charge be nutralized are not ancountered in SDS page. Ultimately protein components separate out only based on molecular weight/ size.