Section A 1. Chemokines are low molecular weight cytokines which induces chemokinesis in leucocytes in inflammation. 2.

To study the gene expression by detection of RNA in a sample 3. Immunodiffusion is a technique to study between antigen and antibodies by observing the formation of precipitation which results from the combination of specifc antigen or antibody wherease immunoelectrophoresis is a method to determine the blood levels of three major immunoglobulin, IgG, IgA, IgM. 4. Agar is a crude agar extracted from seaweed which contain agarose and agaropectin. Agarose is a highly purified which are obtained by removal of agaropectin in agar. Agarose is usually used in immunodiffusion and immunoelectrophoresis. 5. Antiserum contain the whole host (non-specific antibodies) of natural immunoglobulin, while antibody only used to detected specific antigen in a sample. 6. Precipitin ring formed when antigen migrate from the well into the agarose after the reaction between the antibody and antigen. 7. To detecting cytokines or other soluble mediators secreted from a variety of different cell type 8. A cytokine is a small protein released by cells that has a specific effect on the interactions between cells, on communications between cells. 9. Precipitin is an antibody that form precipitation when react with antigen while agglutinin is antibody which causes agglutination. 10. RT-PCR – Reverse Transcription Polymerase chain reaction RIEP – Rocket immunoelectrophoresis Section B 1. There are two types of immunodiffusion, radial immunodiffusion and double immunodiffusion. Both are carried out in a small semisolid medium such as agar. In Radial immunodiffusion, an antigen sample is placed in a well and allowed to diffuse into agar containing a suitable dilution of an antiserum. The antigen will diffuse into the agar. Initially, as the antigen diffuse out of the well, its concentration is relatively high and soluble antigen-antibody adducted are forms. However, as Ag diffuses farther from the well, Ag-Ab complex react with more antibody resulting to the formation of a ring of precipitation, a precipitin ring forms around the well. The area of the precipitin ring is

Various concentration of antigen are loaded in agarose gell’s well. When antigen concentration is increased. each containing single antigenic species allowed to diffuse from separate wells against the antiserum. this indicates the presence of antibody in the antigen of the sera while if more than one precipitin line formed. When precipitin line forms. As equivalence is reached. thereby establishing a concentration gradient. Depending on the similarity between the antigens. . farther the antigen will travel through the gel. a visible line of precipitate. 2. RIEP is a simple. There are two types of immunoelectrophoresis. Spur like results are observed. Three patterns can be observed. Patter of partial identity occurs when the antibodies in the antiserum react more with one of the antigens than the other. different geometrical patterns are produced between the antigen and antiserum wells. By comparing the area of the precipitin ring with standard curve. The two antigens are identical.proportional to the concentration of antigen. rocket immunoelectrophoresis (RIEP) and counter-current electrophoresis. Pattern of NonIdentity occurs when none of the antibodies in the antiserum react with the antigenic determinants. both antigen and antibody diffuse rapidly from wells toward each other. In this test. Higher the amount of antigen loaded in the well. This technique used to check antisera for the presence and specificity of antibodies for a particular antigen. On electr ophoresis. the AgAb complex precipitates and this precipitin line is seen in the form of a rocket. This assay is frequently used for comparing different antigen preparation. When the equivalent point is reached. a precipitin line forms. This shows that the two antigen are immunologically unrelated. this indicate the heterogenicity of the antibody for the antigen in the test sera.the antigen begins to migrate toward anode and interact with the antibody molecules to form a soluble antigen-antibody complex. In Double immunodiffusion. different antigen preparation. Ag-Ab complex form a precipitin line like double immunodiffusion. quick and reproducible method for determining the concentration of antigen in an unknown sample. Pattern of identity occurs when antibodies in the antiserum react with both the antigens resulting in a smooth line of precipitate. the concentration of the antigen sample can be determined. the height of the rocket increases. Counter-current immunoelectrophoresis is a rapid version of double immunodiffusion technique.

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