Exam 1 continued

Question 1

What is a drawback of ANA

Answer 1

lacks diagnostic specificity and needs further serologic tests

Question 2

ANA used in this lab

Answer 2

HEP-2, IgG

Question 3

What can ANA test for

Answer 3

Autoimmune diseases such as SLE (lupus) or rheumatoid arthritis

Question 4

Autoantibodies

Answer 4

some immune cells secrete antibodies that attack cells of the body by targeting the nuclear structures

Question 5

ANA test is an example of

Answer 5

indirect immunofluorescence

Question 6

Serum is added to what and why

Answer 6

added to HEP-2, which has many antigens and is ideal for detection of ANA

Question 7

Positive result for ANA

Answer 7

secondary antibody binds to primary with fluorophore

Question 8

ANA is a

Answer 8

screening test and not a confirmatory test

Question 9

What kind of test is a bradfords assay

Answer 9

colorimetric assay for quantification of protein

Question 10

commassie blue

Answer 10

G250

no protein = red = protonated = cationic/acidic
protein = blue = deprotonated = anionic

results in shifted absorption maximum from 470nm to 595nm when blue

sulfonic groups interact with the amino groups of the protein

Question 11

what do we measure in braford assay

Answer 11

measure optical density (absorbance) vs protein concentration

use reference standard of BSA in dilutions

Question 12

what do we measure in braford assay

Answer 12

measure optical density (absorbance) vs protein concentration

use reference standard of BSA in dilutions

unknown density found, find protein concentration

Question 13

advantages of bradfords assay

Answer 13

interacts quickly, very sensitive, not affected by carbohydrates or ions

Question 14

disadvantages of bradfords

Answer 14

interference with detergents like SDS, dependent on basic amino acid composition (can bind non specifically with AAs)

Question 15

G250 interacts with what part of the protein

Answer 15

carboxylic acid through VDW
Amino groups through electrostatic interaction

Question 16

What is the calibration curve made with in bradfords

Answer 16

BSA

Question 17

Double diffusion

Answer 17

both Ag and Ab diffuse towards eachother, line of precipitation formed when coming into contact

Question 18

double diffusion tests for what

Answer 18

how similar antigens are to one another, identical (forms arc, same epitopes), partially identical, or non identical (bands cross each other without interaction, no common epitope)

Question 18

double diffusion tests for what

Answer 18

how similar antigens are to one another, identical (forms arc, same epitopes), partially identical(arc with a spur), or non identical (bands cross each other without interaction, no common epitope)

Question 19

Radial inmmunodifusion tests for what

Answer 19

finding the optimal concentration relative to antigen/antibodies, or to compare antigens

to quantitate antigens or antibodies, depending on what the solution is and what is added into the wells