Laboratory Quick Guide
Antigen Detection ELISA
Student Workstation Checklist
One workstation serves 4 students.
Item (Label)
Contents
Number
✔
(✔)
Yellow tubes
Violet tube (+)
Student test samples (0.25 ml)
4
❑
Positive control (0.5 ml)
1
Blue tube (–)
❑
Negative control (0.5 ml)
1
❑
Primary antibody (1.5 ml)
1
❑
Orange tube (SA)
Secondary antibody (1.5 ml)
1
❑
Brown tube (SUB)
Enzyme substrate (1.5 ml)
1
❑
2
1
❑
❑
10–20
1
1
❑
❑
❑
12-well microplate strips
50 µl fixed-volume micropipet or
20–200 µl adjustable micropipet
Yellow tips
Disposable plastic transfer pipet
70–80 ml wash buffer in beaker
Phosphate buffered saline
with 0.05% Tween 20
Large stack of paper towels
2
❑
Black marking pen
1
PROTOCOL II
Antigen Detection ELISA
Green tube (PA)
❑
1. Label the yellow tubes with each student’s
initials.
2. Label your 12-well strip. On each strip label
the first 3 wells with a “+” for the positive
controls and the next 3 wells with a “–” for
the negative controls. Label the remaining
wells with your and your partner’s initials
(3 wells each).
Student A
3. Use a fresh pipet tip to transfer 50 µl of the
positive control (+) into the three “+” wells.
4. Use a fresh pipet tip to transfer 50 µl of the
negative control (–) into the three “–” wells.
5. Transfer 50 µl of each of your team’s
samples into the appropriately initialed
three wells, using a fresh pipet tip for each
sample.
6. Wait 5 minutes while the proteins in the
samples bind to the plastic wells.
47
Control
or
sample
Student B
7. WASH:
a. Tip the microplate strip upside
down onto the paper towels, and
gently tap the strip a few times
upside down. Make sure to avoid
splashing samples back into wells.
b. Discard the top paper towel.
c. Use your transfer pipet to fill each
well with wash buffer, taking care
not to spill over into...