Project Monthly Report

In June , the main focus was to familiarize myself with the background for the pass . This required an extensive literary productions search covering the customs of IL-25 in allergic airway inflammation , and the relationship amid mitogen-activated protein kinases (MAPKs ) and bronchial asthma . At the resembling eon , I excessively started to deal familiar with the isolation procedure for B cells and separate techniques that I might recitation in the projectBased on literature freshen up , the concentration of IL-25 utilise to achieve stimulation was in the unspecific range of 50ng /ml to 2 ?g /ml . later on consulting my executive program , I decided to first economic pulmonary tuberculosis 100ng /ml of IL-25 to bear the signalling pathway in perk upd B cells . Then the snip-response of MAPKs (including P38 and P44 /42 ) to IL-25 taste was set up . The time intervals for p38 were 0 , 5 , 15 , 30 minutes and flair hour and the time intervals for p44 /42 were 0 , 1 , 5 , 15 and 30 minutes respectively . Flow cytometry was employ to measure the MAPKs activation using phosphor-specific antibodies to p38 and p44 /42 . No phosphorated p38 was lift up while a slight but not emergence was detected for phosphated p44 /42 . For the second trial , the concentration of IL-25 was same as before , but I set up a positive control by using 10-8M PDB to stimulate MAPKs activation . The time intervals for this trial were 2 minutes and 5 minutes .

A signifi stinkert increas! e in the strike of phophorated p44 /42 was ob work ond , but there was no alternate for IL-25 stimulationSince the two trials did not work well , my supervisor sure me to use CD23 (the marker on the B cells , can serve as a low affinity sense organ for immunoglobulin E and involved in the regulation of immunoglobulin E synthesis , and the CD23 guess to down regulate IL-25 ) to determine the persona of IL-25 in B cells . This set up involved dose-response studies of IL-25 , so B cells were incubated for 24 hours with100ng /ml and 200ng /ml of IL-25 . A epochal change was not observed after staining the cells with anti-human CD23 . I think the sample failed because of my lack of experience in conflate cytometry and staining procedures , so I will replicate this experiment again to get better resultsSince fresh human B cell forwardness involves long time and considering my project timeline , I will use RAMOS B cell line to repeat the above mentioned experiment...If you nece ssity to get a full essay, stage it on our website: BestEssayCheap.com

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