Materials needed for Bradford assay in 542:


Elisa reader(s). Use old at 570, new at 620 nm.

16 P200's (4/bench, 1/pair).

16 P20's (4/bench, 1/pair).

Yellow tips, several hundred.

96 well flat-bottomed plates, 15 (one/pair) with some extras.

50 or so test tubes (for dilution of standard protein).

15 100 ml erlenmeyer flasks or similar containers for dist. water (see below).


Standard protein: Weigh about 25 mg BSA, and dissolve to give
several ml at nominal 10 mg/ml BSA in either isotonic saline (0.9%
NaCl); or PBS with pH 7.0-7.4; plus azide 0.03% (5 mM). Make 10
ml of a 1/10 dilution (nominal 1 mg/ml), and determine the A280
to get an accurate concentration (hydration of the powdered BSA
will generally make the concentration 80-90% of that based on
weight of the powder). The concentration is (A280)/(0.67) mg/ml
for BSA (the denominator is 1.35 for rabbit IgG). Aliquot in 1.5
ml amounts in capped test tubes, one tube/bench.  Label with
the mg/ml based on the A280, e.g. "0.88 mg/ml".

[Each standard curve will use about 1.5 ml BSA at 50 micrograms/ml.
Thus, each bench can be provided with 1 ml BSA at 1 mg/ml, and
each pair can make 2 ml of a 20-fold dilution from this.]

Diluent: distilled water (about 30 ml in a 100 ml erlenmeyer per pair).

Bradford reagent, 3 ml/pair (each pair should consume about 2 ml
for 50 wells). Aliquot in capped test tubes. For 15 pairs, need
45 ml total.

Ethanol in squirt bottles. Make sure there is at least one squirt
bottle/bench, filled, and be prepared to refill them if needed.