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ELISA: Distance Learning SupportSubmission:bolditems are individual; non-bold items can be with a partner-Research on antigens and antibodies (steps 1-2)Antigens are substances on the surface of cells, viruses, fungi, or bacteria that enter thebody from outside. Our immune system works to protect ourselves against theseantigens. When the antigen first enters the body, called the incubation period,macrophage engulfs the antigen and is taken to the T-cell. The helper T-cell shows theantigen to the B-lymphocytes while it fights it. Next, the B-cell clones itself into manyplasma cells that are producing antibodies specific to the antigen. Antibodies areproteins that are produced as a response to antigens. The antigen is defeated with thehelp of the antibodies. Memory B-cells are formed which retain the information about thisparticular antigen and the antibodies needed to fight it.-Notes on ELISA video (step 3)a.ELISA stands for Enzyme Linked ImmunoSorbent Assayb.Procedure:i.Experimental sample with proteins is added to wellsii.Proteins bind to the walls of the welliii.Detergent is used to wash out free proteins and blocks the remainingsurface of the wellsiv.Primary antibody is added which binds to a specific antigenv.Primary antibodies that aren’t bound are washed away using detergentvi.A secondary antibody that has an enzyme-linked colormetric tag is addedto the wells which bind to the primary antibodyvii.Unbound secondary antibodies are washed awayviii.Substrate is added to the wells which binds to the enzyme that is linked tothe secondary antibody which causes a color changeix.This color change indicates that the antigen being tested for is presentc.With the help of ELISA, scientists can make use of the existing antibodies madeby the B-lymphocytes to detect whether they were created for the specificantigen that is being tested.-Diagram of ELISA test (step 4)
-Completion of SRS - Serial Dilutions (steps 7-9)a.1234Tube Dilution1/101/101/101/10Final Dilution1/101/1001/0001/10000b.1234Tube Dilution1/1001/501/251/20Final Dilution1/1001/50001/1250001/2500000c.1234Tube Dilution50/15050/15050/15050/150Final Dilution1/31/91/271/81The first tube has a concentration of 200 ng sample/mlSo, the second tube has a concentration of 200 x= 200/3 ng sample/mlSo, the third tube has a concentration of 200/3 x= 200/9 ng sample/mlSo, the fourth tube has a concentration of 200/9 x= 200/27 ng sample/ml-Omit steps 12-13-Notes on video (step 14)a.

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