Molecular Basis of Disease Lab Report Example | Topics and Well Written Essays - 1500 words

Molecular Basis of Disease - Lab melodic theme ExamplePeople lacking the absorption ability of vitamin B12 remain with the disease for the rest of their lives. Parietal cells constitute in the gastric cells are responsible for production of hydrochloric acid which is important in digestion as it provides a favorable medium for enzyme reaction. The serum found from patients having this disease react with ? and ? sub units. The antibodies are utilize for diagnostic purposes. This experiment involves testing of serum of antibodies from various patients to find out if they react with the sodium pump which contains ? and ? sub units. This get out help in diagnosing pernicious anemia. A grovel is used in this experiment since its abdominal cavity structure is similar to that of a man. A primary antibody and a secondary antibody are also used in this experiment as they react with the mouse antigens. Anti proton pump of a human beingness can also cross react with proteins of the mou se making the mouse a favorable warning for the experiment. From the experiment we expect antibody response to help us diagnose pernicious anemia. Aims The aim of this experiment is to determine whether samples from patients contain antibodies and also to diagnose patients having the disease through Western blotting Immuno histochemistry using the gastric proton pump. Materials and Methods The materials used allows SDS polyacrylamide gel (80%, resolving gel 40% upper stacking gel), jelly running buffer (25 mM Tris, pH 5.3, 0.192M Glycerin, 0.1% SDS), Gel apparatus, power supply, mouse stomach protein,5? SDS sample buffer (0.315 Tris, pH 6.8, 25% glycerol 10% SDS, 5% 2-beta mercapto neutral spirits, 0.025% Bromophenol blue), Protein MW standards, heat block, Gel lode tips, transfer apparatus, transfer buffer (25mM Tris, 192mM glycine,20% methanol), nitrocellulose membrane, filter paper, 0.1% Ponceau in1% acetic acid, 0.1M NaoH, TBS Tris buffered saline, block solution TBS conta ining 5% skim milk powder. Materials required to obtain a mouse stomach include a slide containing section of mouse stomach, xylene, hemoglobin, acid alcohol, Scotts hit water, eosin, DPX mounting media and stew slips. purpose to obtain mouse stomach morphology The slide containing mouse stomach is incubated in xylene and ethanol for two minutes respectively. After this the slide is rinsed in tap water for 30 seconds. The slide is thence incubated in the hemoglobin for 2 minutes. The slide is rinsed again in tap water for 30 seconds by and by which it is placed in 1% acid alcohol for 3 seconds after which it is rinsed again in tap water for 30 seconds. The slide is incubated in Scotts tap water for 30 seconds after which it is swashed in water for 30 seconds. The slide is then placed in eosin for 4minutes and excess is blotted off by a paper towel. The slide is then placed in 80% ethanol and again 90% after which the slide is incubated in ethanol for 2 minutes then allowed to ai r dry. A expel of DPX mounting medium is placed on the section and then covered by a cover slip. An observation is made under the microscope and diagrams made as follows. Fig 1 showing a mouse stomach with Haematoxylin To prepare the stomach protein sample, 200l of protein is placed into a microfuge tube and 50l 5? SDS sample buffer is added. The marker and the protein sample tubes are spine for a few seconds to learn the liquid to the bottom of the tube. The MW markers and proteins are then loaded using a special gel loading tips. The apparatus is then connected to a power supply of 200V and run for 1 hour until