ELISA Enzyme-linked Immunosorbent Assays (ELISA)
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Questions and Answers

What is the purpose of the blocking buffer/solution in a Direct ELISA?

  • To promote the binding of the primary antibody to the antigen
  • To facilitate the binding of the antigen to the well surface
  • To prevent non-specific binding of antibodies to the well surface (correct)
  • To enhance the hydrophobic interactions between proteins and the well surface

    • In Indirect ELISA, which molecule binds to the primary antibody?

  • The capture antibody
  • The secondary antibody (correct)
  • The blocking buffer/solution
  • The antigen

    • What is the role of the capture antibody in Sandwich ELISA?

  • To bind to the primary antibody
  • To bind to the antigen in a specific manner (correct)
  • To prevent non-specific adsorption of proteins to the well surface
  • To enhance the hydrophobic interactions between proteins and the well surface

    • Why are microplates used for ELISA made of polystyrene?

    <p>Polystyrene is hydrophobic, allowing passive adsorption of biomolecules through hydrophobic interactions</p> Signup and view all the answers

    What is the function of a mature naive B cell during the initial encounter with a pathogen?

    <p>Act as an antigen-presenting cell</p> Signup and view all the answers

    What happens to the B cell after a helper T cell specific for the same antigen activates it?

    <p>It differentiates into memory B cells and plasma cells</p> Signup and view all the answers

    What is the function of memory B cells?

    <p>Secrete antibodies to fight against the pathogen</p> Signup and view all the answers

    What is the purpose of using an enzyme-linked anti-mouse 2˚ antibody in the given scenario?

    <p>To detect different antigens with a single antibody</p> Signup and view all the answers

    Which region of the antibody does the enzyme-linked anti-mouse 2˚ antibody bind to?

    <p>Constant region</p> Signup and view all the answers

    What is the function of the control line in the lateral flow immunoassay?

    <p>To indicate if the result is negative or positive</p> Signup and view all the answers

    What is the purpose of the chicken IgY antibodies in the lateral flow immunoassay?

    <p>To bind to the tagged antibodies in the conjugation pad</p> Signup and view all the answers

    Which type of antibodies can be used for research as mentioned in the text?

    <p>Monoclonal or polyclonal antibodies</p> Signup and view all the answers

    How do antibody-bound antigens reach the test line in the lateral flow immunoassay?

    <p>By binding to another antigen-specific antibody on the test line</p> Signup and view all the answers

    Study Notes

    Direct and Indirect ELISA

    • Blocking buffer in Direct ELISA prevents non-specific binding of antibodies to the plate.
    • In Indirect ELISA, secondary antibodies bind to the primary antibodies enhancing signal detection.

    Sandwich ELISA

    • The capture antibody attaches to the target antigen, providing specificity and allowing for detection.

    Microplates

    • Polystyrene microplates are used for ELISA due to their high protein-binding capacity and compatibility with various assays.

    Immune Response

    • Mature naive B cells recognize and bind to specific antigens during initial encounters with pathogens, beginning the immune response.
    • After activation by helper T cells specific to the same antigen, B cells proliferate and differentiate into antibody-secreting plasma cells or memory B cells.

    Memory B Cells

    • Memory B cells persist long-term after an initial immune response, providing a quicker and stronger response upon subsequent infections.

    Antibody Detection

    • An enzyme-linked anti-mouse secondary antibody is used to amplify the detection signal in ELISA and binds to the Fc region of the primary antibody.

    Lateral Flow Immunoassay

    • The control line confirms the validity of the test by showing that the assay worked properly.
    • Chicken IgY antibodies serve as a detection agent in lateral flow assays due to their unique properties and reactivity.

    Antibody Research

    • Different types of antibodies, including polyclonal and monoclonal antibodies, are utilized for research applications.

    Antigen Mobility

    • Antibody-bound antigens move to the test line in a lateral flow immunoassay through capillary action, allowing for visualization of results.

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    ELISA Enzyme-linked Immunosorbent Assays (ELISA) - Activity 3 Slides PDF

    Description

    This quiz covers the method for detecting and quantifying a specific protein in a protein solution using ELISA. It also includes information about the antibody-antigen binding and the non-specific adsorption of the antigen or antibody in Immunosorbent Assays.

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