Bio-Rad Comparative Proteomics Kit II: Western Blot Module - Manual

25
Quick Guide
Q
UICK GUIDE
1ml
Comparative Proteomics Kit II:
Western Blot Module – Quick
Guide
Lesson 1 Quick Guide
1 Label one 1.5 ml fliptop micro tube for each of
five fish samples. Also label one screw-cap micro
tube for each fish sample.
2. Add 250 µl of Bio-Rad Laemmli sample buffer to
each labeled fliptop microtube.
3. Cut a piece of each fish muscle about 0.25 x 0.25
x 0.25 cm 3 ( ) and transfer each piece into a
labeled fliptop micro test tube. Close the lids.
4. Flick the microtubes 15 times to agitate the tissue
in the sample buffer.
5. Incubate for 5 minutes at room temperature.
6. Carefully transfer the buffer by pouring from each
fliptop microtube into a labeled screw-cap
microtube. Do not transfer the fish!
7. Heat the fish samples in screw-cap microtubes
for 5 minutes at 95°C.
Lesson 2 Quick Guide
1. Set up Mini-PROTEAN 3 gel box.
2. Prepare a Ready Gel cassette by cutting
along the black line on the bottom of the
cassette with a razor blade and pulling off
the plastic strip, as indicated on gel
cassette.
3. Remove the comb from the Ready Gel
cassette.
4. Place Ready Gel cassette into the
electrode assembly with the short plate
facing inward. Place a buffer dam or
another Ready Gel cassette on the
opposite side of the electrode assembly,
with notch on buffer dam facing inward.