Answered: 1. What term is the word “antigen”…

Human Anatomy & Physiology (11th Edition)

11th Edition

ISBN:9780134580999

Author:Elaine N. Marieb, Katja N. Hoehn

Publisher:Elaine N. Marieb, Katja N. Hoehn

Chapter1: The Human Body: An Orientation

Section: Chapter Questions

Problem 1RQ: The correct sequence of levels forming the structural hierarchy is A. (a) organ, organ system,...

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Can you please check and answer the questions that not answered, that is not graded https://www.studocu.com/en-us/ document/university-of-arkansas/microbiology/other/immunology-lab-worksheet-student/7960193/view Questions from ELISA Simulation Introduction Document (posted in Lab Module 8 folder): 1. What term is the word “antigen” derived from? 2. Where are antibodies found in the body? Plasma of the blood 3. In general, what is the cause of an autoimmune disease? 4. What kind of antibodies are present in patients with systemic lupus erythematosus (SLE)? 5. What does ELISA stand for? enzyme-linked immunosorbent assay 6. In general, what is an ELISA test used to detect? to measure the concentration of antibodies or antigens 7. In an indirect ELISA, which antibody is linked to the enzyme? (i.e the primary antibody from the blood serum or the secondary anti-human antibody) Questions from HHMI Virtual Immunology Lab Website Diagnosis (The answers to the following question can be found in the “Diagnosis” tab) 8. What are the 3 important limitations of an ELISA? Explain each. a.A positive result:confirming a presence of an antibody but it not necessarily making the patient sick. b.A false negative:where the amount of antibodies is too low to be measured. c.A positive result:may occur if an unrelated antibody reacts with the antigen nonspecifically. Background (The answers to the following questions can be found in the “Background” tab) 9. What determines if a patient has an infectious or autoimmune disease? the glisa test 10. What does a positive result indicate? the antibody is there and implies that the person has encountered a particular disease. 11. The watery fluid of the blood is called ______serum_______. 12. What is allowed to react with the target antigen? a portion of the serum containing the antibody 13. Detection becomes possible when _________a second antibody is added____________________________________. 14. What is the secondary antibody prepared from? 15. Once isolated, the secondary antibody can be____ chemically linked to a system that can produce a detectable signal_______________________________. 16. What does the signaling system consist of? consists of an enzyme attached to the second antibody 17. What happens when the appropriate chemical (substrate) is added? the enzyme converts to a colored substance that can be measured. 18. The ELISA test directly quantifies how much _________________ is present by the amount of _________ produced. 19. How does the amount of color produced in the ELISA relate to the amount of antibody found in the patient (the primary/first antibody)? 20. From Figure 1 (click on it), what are the 4 steps of an ELISA protocol? a.Bind to sample support b. Add primary / wash c. Add secondary antibody enzyme conjugate d.Add substrate Lab Notebook Click on the Notebook tab and begin the lab itself. (Click “start over” to begin the lab if this option appears.) Proceed through the entire lab simulation protocol. Be sure to read the captions below the pictures (left side) and the information in the lab notebook (right side). 21. Which disease are you testing the three patients for? (Click on the disease name to read more about it) 22. Why are you using the centrifuge in step 1? What will you obtain? precipitate the blood cells and obtain the serum 23. What are you preparing in Step 2? preparing serial dilutions. 3 solutions to make diff dilutions 24. What does PBS stand for? Why is it used in Step 2? 25. (Step 3) What has the ELISA plate been pretreated with? Why? With sle antigen to get the antigens to bind to the plate. 26. (Step 4) What is the positive control? the anti-dna primary antibody 27. What is a primary antibody? (define) the second antibody used in a immune assay that affects the pr 28. (Step 4) What is the negative control? buffer 29. (Step 4) Why is it necessary to have a positive and a negative control? the elisa isnt always conducted under appropriate condition so if either test gives unexpected results the assay cant be trusted 30. (Step 5) Why is the plate incubated? Why is incubation done at 37ºC? ensures that the antibodi present in the sample will interact correctly with the antigen 31. (Step 6) Why wash the plate? helps remove any antibodies that dont react with the sle antigens 32. (Step 7) What is a secondary antibody? (Click on the definition) an antigen from a diff species which is used in an immunoassay that detects primary antibody 33. (Step 7) What is the attached enzyme in this assay? hrp enzyme 34. (Step 7) What is the specific substrate for HRP? What color does it produce? substate abts, produces a yellow solution 35. (Step 10, in “why”) How can the yellow color be quantitatively measured? At what wavelength? can be estimated by the eye or 4/4 nanometers. 36. Results (Indicate on this page and on the computer which boxes turned colors.) A B C + (positive) - (negative) 1:2 Yellow Yellow Yellow 1:10 yellow yellow yellow 1:100 yellow yellow 37. Did you complete the ELISA correctly?Yes If yes, proceed to #38 If no, proceed to #39 38. What do the results indicate about: Patient A ________likely to have sle _________ Patient B ______Might have it but further testing is required ___________ Patient C ______Does not have it ___________ 39. Explain what you did wrong and what you will need to do next time. Did your incorrect procedure provide you any results?

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