476717
Description
Flashcards by sophietevans, updated more than 1 year ago
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Created by sophietevans
almost 11 years ago
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| Question | Answer |
| What were the pellet and supernatant used for in this/these assay(s)? | The pellet was used to determine the equivalence point of BSA-anti-BSA and therefore the BSA concentration, whereas the supernatant was used to detect the excess, to prove in an inverse manner that the equivalence point was accurate. |
| Why does excess antigen not result in maximal precipitation of antibody and antigen? | With excess antigen, the complex is not precipitated as both the binding sites of the antibody will be saturated in excess so cannot link to antigen on other antibodies. |
| What is the quantitative precipitin curve used to determine? | The concentration of the antigen or antibody (whichever is not known and is being sought in a sample). |
| What two things need to be known in order to calculate the concentration of antigen/antibody in a quantitative precipitin curve? | The ratio of antigen:antibody and the concentration of the antibody/antigen that is known. |
| What piece of equipment was used to establish the level of precipitation at the different antigen concentrations? | A spectrophotometer. |
| What would you expect the Ouchterlony slides for BSA and anti-BSA to show either side of the equivalence point? | BSA: When the antigen is at a low concentration, no precipitin would be seen on the slide as all of the small amount of precipitated complexes would have been captured in the pellet. After the equivalence point, when the antigen is in excess, a continuous curve of precipitin would be seen as the excess BSA would have been caught in the supernatant, and the wells would have identity...because they're identical. Anti-BSA: Before the equivalence point, a continuous precipitin curve would be formed as there would not be enough antigen for all the anti-BSA to be complexed. The excess would be caught in the supernatant and complexed in the Ouchterlony slide. After the equivalence point, when the antigen is in excess, no precipitin would be present as all of the antibody would have precipitated and been captured in the pellet. |
| What type of reaction/technique is this? | A quantitative precipitation reaction. |
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