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Fig. 34. The pH gradient profile indicated by the Broad pI calibration kit for the gel
shown in Fig. 33. The gel was projected onto a 25 x 25 cm format for band
measurement. The proteins, starting from the cathode and their corresponding pI´s are:
lentil lectin (basic) 8.65
lentil lectin (middle) 8.45
lentil lectin (acidic) 8.15
horse myoglobin (basic) 7.35
horse myoglobin (acidic) 6.85
human carbonic anhydrase B 6.55
bovine carbonic anhydrase B 5.85
ß-lactoglobulin A 5.20
soybean trypsin inhibitor 4.55
amyloglucosidase 3.50
Molecular weight measurement
Molecular weights of globular and SDS-denatured proteins are easily
measured using PhastGel homogeneous or gradient media and one of
the
GE Healthcare
calibration kits. Pour calibration kits are
suitable for use with PhastGel media; the high molecular weight
(HMW) kit for native or SDS-denatured proteins, the low molecular
weight (LMW) kit for SDS-denatured proteins, a high molecular
weight calibration kit especially prepared for SDS runs with
PhastSystem (HMW-SDS) and a molecular weight kit intended for
SDS runs of very small proteins or peptides (PMW).
GE Healthcare
calibration kit proteins come in convenient
freeze dried mixtures. The proteins are exactly characterized and
highly purified. Exact protein amounts per vial have been chosen to
give bands of equal intensity on staining with Coomassie. Table 2
below shows the molecular weight ranges covered by the calibration
kits.
6. Evaluation and presentation of data