Download Immunodiagnostics: A Practical Approach (Practical Approach by R. Edwards (Editor) PDF

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By R. Edwards (Editor)

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Extra info for Immunodiagnostics: A Practical Approach (Practical Approach Series)

Example text

8. Repeat this cycle twice more. 9. 4 and store at 4°C in same phosphate buffer. B. Affinity chromatography 1. Absorb specific antibodies onto gel by mixing with antiserum, volume depending on litre, approximately 10 ml per g. Mix over 1-2 days. 2. 15 M sodium chloride, at approximately 20 ml per h until OD at 280 nm has returned to baseline. 3. 0 containing 20% acetonitrile. 0 (adjust pH with acetic acid) containing 20% acetonitrile. 0 (adjust pH with acetic acid) containing 20% acetonitrile. 1 ml).

Same proportion of different forms. 3. The substance used as standard should be chemically pure. 32 1: Principles of immunodiagnostic tests and their development 4. g. , should be similar for both standards and sample. 5. e. reference immunoassay and not a bioassay. Some preparations may give quite different potencies in bioassays compared to immunoassays. 6. Standards must be produced in a form which ensures maximum stability. The chemical identity and degree of purity of the material selected for use as standard has to remain a matter of judgement.

Mix until dissolved. 5. Add papain corresponding to 16% of the IgG (w/w). Gently mix until dissolved. 6. 6 mg/ml. 7. Shake the solution at 37°C for one hour and then allow to stand at 37 °C for a further 23 hours. 8. 8 mg/ml and shake for 15 minutes at 37°C. 9. Centrifuge (4500 rpm for 45 min) to isolate the precipitate (Fc). Decant the supernatant which contains Fab. 10. 9% NaCI). 11. For further purification affinity chromatography (with immobilized specific antigen) or ion-exchange may be employed.

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