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  Antigen Antibody Reactions

Antigen Antibody Reactions Immuno Fluorescence:

Fluorescence:

Fluorescence is a phenomenon stated as an emission of light of a specific color by a substance whenever it is irradiated with light of a different color.

Fluorochromes:

Substances that exhibit the phenomenon of fluorescence are termed as fluorochromes. Fluorochromes are organic substances acquired by synthesis or natural techniques.

There are lots of fluorochromes: For labeling antibodies the Fluorochrome must have the properties shown below:

1. Must have chemical group capable of making stable compounds with protein molecules (i.e., azo carbamide, sulfonamide, thiocarbamide,)

2. Fluorochromes which have not been coupled must be easy to eliminate.

3. Fluorescent light emitted must be influential & must refuse minimally.

4. Must not have chemical groups which in combination with a protein could disturb the reaction.

5. The fluorescent color of the conjugate must be dissimilar from that given off by normal cellular substrate (i.e., Autofluorescence)

6. The conjugate resultant from the coupling with the protein must be as stable as possible in normal situation of preservation.

7. Immunological reactivity must be retained.

8. Coupling technique must be simple

9. Fluorochrome must be easy to synthesize, easy to manipulate, economical and safe to those who manipulate them.

 

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