The Allergens for the Cows Milk - Essay Example

The subjects should be exposed to raw milk and baked milk and the immediate allergenic response must be detected using the ELISA kits for casein and beta-lactoglobulin. The total duration for the test is 45 minutes only. (oxoid.com, 2009). These kits have a greater sensitivity for the detection of allergens.  If food materials are used for the diagnosis, then proper care must be taken to dilute and centrifuge the samples such that the supernatant will contain the material to be tested. Usually, food materials are weighed and crushed using the pestle and mortar. A suitable is added to the raw material and centrifuged at high speed such that all the tiny particulates settled down and only the liquid substance is present at the top. 100 microliters of the sample are enough for the detection of the allergen. (oxoid.com, 2009).
            The sera of the subjects are prepared, and this is kept in the Eppendorf, 100 microliters of the serais transferred to the antibody-coated wells. 100 microliters of the standards are also added to the wells and mixed for 10 seconds gently and incubated for 15 minutes. After this, the liquid in the well is dumped off and the wells are washed five times using the wash buffer. The wells are then made sure that no buffer sticks to its them tapping it on the absorbent paper towel. Then the conjugate solution is added to each well. 100 microliters can be added to each well. (elisas.com, 2011). The plate is then gently mixed for 10 seconds and then again incubated for 15 minutes. The liquid in the wells is dumped again. Then using the wash buffer the wells are thoroughly washed five times to ensure that the conjugate that has not bound to the well is removed completely.  On the absorbent paper towel, the wells are tapped firmly before the next step is continued.  Finally, the substrate solution at a definite concentration is added to the wells. 100 microliters are added to the wells and mixed gently and incubated again for 10 minutes. The stop solution (100 microliters) is finally added to each well and again mixed gently for 10 minutes. (elisas.com, 2011). The results are observed visually, and the concentration can be identified by using a microtiter reader.  For these tests, the results must be read within 30 minutes. These kits can detect the milk protein concentrations by parts per million. The controls used for this test are usually skimmed milk powder.  (elisas.com, 2011).

Thus, we can investigate the concentration of the allergen in the sequential addition of the milk produced in the subjects under investigation.

Required Chemicals and Materials:

• ELISA KITS ( case in detection kit and Betalactoglobuline Kit).
• • Microtiter plates
  • Skimmed milk powder
  • Baked milk food products
  • Raw milk
  • Sera of the subjects
  • Sera of the control subjects
  • ELISA plate reader.

  SCHEDULE

   

  DATES AND NO. OF TEACHING WEEKS

   

  Planning stage

  Researching topic, using relevant textbooks and journals.

   TW 1 & 2

  Write-up of literature review

  TW 3 - 6

  Gather information on methods to be used in experiments

  TW 3 - 6

  Find out what chemicals need to be ordered

  TW 3 - 6

  Complete COSHH safety forms

  TW 3 - 6

  Submit plan of investigation to include a timetable for data collection

  By 5 pm on Monday 14th November 2011

  Receive feedback- discuss progression and agree a start date with supervisor

  By Friday 25th November

   

  Data collection and writing up stages

  Instrument training

  Monday 28th November

  Collection of specimens

  TW 10

  Lab-based work

  TW 10 - 12

  Interpreting data

  Semester 2 TW 1 - 2

  Write up a scientific paper

  Semester 2 TW 6 until enrichment week

  Prepare for oral presentation

  Semester 2 TW 7 onwards

   

  Statistical analysis:
              Sample T-test and ANOVA test are used for the statistical analysis. Bar charts and graphs are used at suitable places to demonstrate the differences in the allergic responses of cow’s milk and baked milk.