IMMUNOLOGY

INMUNOLOGÍA


4. IMMUNOLOGY

INTRODUCTION TO ELISA REACTION

Your students will learn the basic principles of the Enzyme-linked Immunosorbent Assay (ELISA) in this precise and sensitive antibody-based detection kit.

Experiment components do not contain human serum.

> Kit includes:
Antigens, primary and secondary antibodies, peroxide co-substrate, hydrogen peroxide, ABTS substrate, phosphate buffered saline, tubes, plates, and transfer pipettes. 
> You need:
Distilled water, 37º C incubation oven, automatic micropipettes with tips and laboratory glassware.

icons-ref-greenELISA1
      10 groups of students

imgcat--ELISA

SIMULATION OF HIV DETECTION BY ELISA

This experiment’s aim is to provide students the molecular biology and pathogenesis basics of acquired immunodeficiency syndrome (AIDS).

An HIV test detects HIV infection indirectly using an ELISA test against HIV antibodies in the blood. The test works by taking antibodies from the patient’s blood and adding them to a microtiter plate coated with HIV antigen. If HIV antibodies are present in the blood, they will bind to the antigens on the plate. This binding is detected with an enzyme-linked secondary antibody that causes a color change upon addition of substrate. In this experiment, students will perform an ELISA test by coating microtiter plate wells with simulated HIV antigen and then test simulated donor serum for anti-HIV antibodies.

> Kit includes:
Simulated HIV antigens, serum samples, antibodies, buffers, microtiter plates, assorted pipettes and microtest tubes. 
> You need:
37ºC incubation oven, micropipettes with tips, pipette pumps, laboratory glassware, and distilled water.

icons-ref-greenELISA2
     10 groups of students

imgcat--ELISAhvi

QUANTITATIVE ELISA

Antibodies are highly specific in their recognition of antigens. This ELISA experiment demonstrates the quantitation of varying concentrations of viral antigens as detected by the intensity of the color reaction due to the accumulation of products.

> Kit includes:
Antigens, primary and secondary antibodies, substrate solution, phosphate buffered saline, blocking agent, stop solution, tubes, plates, and transfer pipettes.
> You need:
Distilled water, 37º C incubation oven, micropipettes with tips, laboratory glassware.

icons-ref-greenELISA3
     6 groups of students

SIMULATION OF HIV DETECTION BY WESTERN BLOT

The second assay used to confirm a positive HIV ELISA result is the Western Blot. Students separate protein samples from hypothetical patients on agarose gels, transfer the samples to a membrane and detect the simulated HIV proteins. This kit is an introductory level experiment.

> Kit includes:
Samples, standard molecular weight markers, protein agarose, various buffers and reagents, PVDF membrane, filter paper, stain, 1 ml pipette, 100 ml graduated cylinder.
> You need:
Electrophoresis apparatus, power supply, micropipettes with tips, microwave, incubation oven, shaker platform, lab glassware, small plastic trays, microtest tubes, pipette pumps, metric rulers, distilled water, methanol, and glacial acetic acid.

icons-ref-greenELISA4
     6 groups of students

imgcat--WESTERN-BLOT

WHAT’S IN MY LUNCH? QUANTITATIVE MILK ALLERGY ELISA

The second assay used to confirm a positive HIV ELISA result is the Western Blot. Students separate protein samples from hypothetical patients on agarose gels, transfer the samples to a membrane and detect the simulated HIV proteins. This kit is an introductory level experiment.

> Kit includes:
Dilution buffer, wash buffer, whey antigen, primary and secondary antibody, aminosalicylic acid, hydrogen peroxide, microtiter plates, microcentrifuge tubes, 15 and 50ml conic tubes and transfer pipettes.
> You need:
Dilution Various food samples, distilled water, flasks, paper towels, 37ºC incubator, disposables lab gloves, safety goggles, automatic micropipettes and tipes, digital camera (or cell phone with camera), computers with internet and image analysis program.

icons-ref-greenELISA5
     10 groups of students

266_Gallery

RADIAL IMMUNODIFFUSION

Radial Immunodiffusion quantitatively determines the level of an antigen. Antibody is incorporated into liquefied agar and allowed to gel. The antigen is added to small wells and radiates throughout the antibody-containing medium, leaving a precipitate throughout the gel. The amount of diffusion is quantified.

>Kit includes:
Antigen and antibody, petri plates, pipettes, well cutters, agarose, buffer and microtest tubes.
> You need:
Automatic micropipettes with tips, waterbath, microwave or hot plate, incubation oven, laboratory glassware, pipet pumps or bulbs, rulers, paper towels and distilled water.

icons-ref-greenINMUNO1
     10 groups of students

IMMUNOELECTROPHORESIS

Learn how Immunoelectrophoresis identifies proteins based on their combined electrophoretic and immunological properties. This method is useful to monitor antigen and antigen-antibody purity and to identify a single antigen a mixture of antigens. In this experiment, serum proteins are separated by agarose gel electrophoresis and the point of equivalence is observed by the antigen-antibody complex formation.

> Kit includes:
Proteins, antibodies, reagents, agarose, buffer, transfer pipettes, well cutters and paper wicks.
> You need:
Horizontal electrophoresis apparatus, power supply, automatic micropippetes with tips, microwave or hot plate, incubation oven, laboratory glassware, microscope slides, paper towels and distilled water.  

icons-ref-greenINMUNO2
     10 groups of students

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