Skip to main content

Network theory and it`s experimental evidence|clona?l selection theory?

     
          Network theory and it`s experimental evidence

-        Any discussion regarding of immune response not come to the end without mentioning Jern`s idiotype network theory.
-        In 1973, Neil Jerne proposed this theory so to give possible solution for regulation of immune system in 1985 he was honored with Nobel prize for this work.
-        This theory suggest that antigen enter the inside the body and activate very specific done of B cell secreting antibody against it.
-        The antigen binding site of an antibody composed of overlap between light and heavy chain domain called as idiotype.
      
 -        This recognizing are said to be complementary to antigen.
-        Antibody molecule itself being a protein so can behave as an antigen.
-        Antibody molecule against idiotype which exhibit internal image of an external antigen.
-        Likewise, anti-ant idiotype antibody as external images of primary antibody.
-        Theoretically this sequence can go further consisting network od idiotype antibodies, however experientially it has been observed that is extend to just their steps.
-        As a consequence of such antibody network antigen is readily remove from circulation.
-          At the same time each antibody feedback regulates formation of antibody once step ahead. Thus, controlling the overproduction of antibody

Experimental evidence: -
            Hemolytic plaque assay (jern`s hemolytic assay)
1.    This assay is used to quality number of antibodies producing plasma cell.
2.    Assay was introduced by jern`s and modified by qunning in 1968.

Protocol: -
Ø First mouse is immobilized with sheep RBCs
Ø After a fixed incubation period the spleen from mouse isolated.
Ø Then the solenocytes from the spleen are separated which contain plasma cell and this plasma cell produce antibody against sheep`s RBC`s.
Ø A microchamber of slides is prepared using two slides.
Ø The solenocytes are introduced in the microchamber along with sheep RBC`s and complement protein.
Ø The microchamber is incubated at 370c for 2-3 hours.
Ø During this incubation period, plasma cell secretes the antibody against RBS`s (of IgM type).
Ø The secreted antibodies then bind to sheep RBC`s present in surrounding.
Ø Once antibody binds to sheep RBC`s present in surrounding.
Ø Once antibody binds to sheep RBC`s complement lysis the RBC`s.
Ø Due to this lysis of RBC`s a clear zone is produced around the antibody producing cell.
Ø Number of plaques are directly proportion to number of antibodies producing plasma cell.

         

Ø  
Ø  

Comments

Popular posts from this blog

Yeast two hybrid assay ?

Yeast two hybrid assay: -                                          Study interaction, detecting protein-protein interaction.   Principle of the two-hybrid: - A] Standard model of transcription activator: -     - The DNA binding domain of an activator binds to an enhancer.     - The activating domain interacted with the based complex. Recruiting it the promoter       B] Two hybrid assay for protein-protein interaction using gene clone technique linked the gene one protein to the part of a gene encoding a DNA binding domain, to encode 1 hybrid protein, linked the gene for another protein to the part of the gene encoding a transcription activating domain to encode a two hybrid proteins are introduce into yeast...

What is random priming?|What is non radioactive Labelling?|What are random primers used for?

                          Random primer labelling (primer extension) Ø Gene probes, clone or TCR amplified and oligonucleotide probes can be random prime labelled with radioactive isotopes and non-radioactive labelled DIG Ø Random prime labelling of DNA fragment (double or single stranded DNA was developed by Fein Berg and Bolgesteinds an alternative to nicker translation to produce uniformly labelled probes. Ø Double stranded DNA is degenerated and aneled with random oligonucleotide primer. Ø The oligonucleotide primer too 5`―› 3` polymerase which synthesis labelled probe in presence of labelled nucleotide pressure. PCR DIG labelling: -         DIG DUTP is incorporated during PCR cycle into the DNA strands amplified from the DNA target. Photo biotin labelling: - Photo biotin labelling is a chemical reaction not an enzymatic one. ...

Detection of DNA binding ?

                                                Detection of DNA binding Radioisotope: - ·        Nucleic acid probe can be labelled using radioactive isotopes. ·        Most common process but less popular today become of safety consideration as well as coast and disposal of radio waste product. ·        However, radio labelled probes are the sensitive as they provide the highest degree of resolution currently available in hybridization assay. Non-radioactive labelled: - ·        Compared to radioactive labelled use of non-radioactive labelled have several advantages 1.     Safety. 2.     Highest stability of probe. 3.     Efficiency ...