A capillary tube of whole blood is collected and centrifuged to obtain a plasma sample. A 1 in 4 dilution of the plasma is made and analysed using a cholesterol assay like the one you performed in the lab. 10 μL of both the diluted plasma sample and a cholesterol standard are assayed using 1 mL of cholesterol reagent. Following incubation at 37 °C the samples are measured at 500 nm. The experiment yields the following data: A500 standard: 0.22 A500 sample: 0.59 Concentration standard: 3.5 mM What is the concentration of total cholesterol in the diltuted plasma (only type in your numerical value in the units mM to 2 decimal places)?
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- Explain why a semi-log plot should be used for determining antigen concentration by ELISA.Which of the following describes the proper order for a competitive ELISA? first coat wells with test sample, and then add enzyme conjugated detection antibody. Firaly, add subsrate first coat wells with antigen, and then, incubate test sample with capture antibody. Next, add incubated sample to plate. Finally, add substrate first coat wells with capture antibody, and then, add test sample. Next, add enzyme-conjugated O detection antibody, and finally, add substrate first coat wells with antigen, and then, add primary antibody. Next, add enzyme-conjugated detection antibody, and finally, add substrateWhich of the following best describes a positive reaction in an ELISA? A color change takes place. All of the molecules in the ELISA coagulate. There is a pH change. If the color of the ELISA does NOT change, the result is positive.
- You perform an ELISA to test hospital soap samples for contamination with Pseudomonas aeruginosa. Other than the different molecule of interest, this ELISA works exactly like the one you saw in the video. You run the soap sample, a negative control, and a positive control, all in duplicate. How would you interpret the following results? Laboratory Notebook: ELISA – Pseudomonas aeruginosa Well Contents Observation at 15 min Result 1 Negative control 2 Soap sample 3 Positive control 4 Negative control 5 Soap sample 6 Positive control 7 Empty well 8 Empty well The soap is POSITIVE for the Pseudomonas aeruginosa antigen. You cannot interpret the soap data, because the NEGATIVE control didn’t work. The soap is NEGATIVE for the Pseudomonas aeruginosa antigen. You cannot interpret the soap data, because the POSITIVE control didn’t work.1. How do you determine the concentration and dilution of a solution using the tube dilution method? 2. Decide on what to do with this scenario. You are required to measure 1 mL of serum to be added to 5 mL reagent in a clinical chemistry assay. However, the amount of specimen that is left for you to work with is only 0.5 mL. What will you do if you are not allowed to take another specimen from the patient anymore?What is this type of test called? What specific antigen did we use it for? What organism was identified with the test? How did you know it was positive? How did you know it was negative? How did you know the test was valid?
- The stool samples submitted by the two friends were enumerated following enrichment. The technician prepared serial dilutions. A volume of 1 mL of stool was mixed with 1 mL of diluent. From this dilution, they then took 1 mL and added it into 1 mL of diluent. The operation was repeated 5 more times. After finishing the serial dilution, 100 μ�L has been spread on different TSA plates. Following 24h incubation, the technician counted 23 colonies at the dilution 4. What is the bacterial concentration in the enriched culture? [1 mark]You are tasked with measuring the quantity of prostate specific antigen (PSA) in a urine sample using an ELISA strategy. Describe and schematically depict how you would perform the test given the following materials: anti-PSA, biotinylated anti-PSA, streptavidin-HRP, HRP substrate kit (HRP = horse radish peroxidase).Which tube will serve as the control in the “Normal Serum Bactericidins” portion of this lab activity? Tube A (Unheated normal serum) Tube B (Normal serum heated at 56 degrees C for 30 minutes) Both Tubes A( Unheated normal serum) and B (Normal serum heated 56 degrees C for 30 minutes) Tube C (Saline) There is no control in this activity.