QUESTION 1
You have designed a new protocol for isolating Golgi apparatus from cell suspension culture using a
combination of sucrose step gradient and free flow electrophoresis (FFE, refer Figure 1 for the
strategy). In order to convince your colleagues that FFE produces highly enriched Golgi fraction
suitable for downstream sensitive applications, you first collected fractions from FFE, followed by
protein quantitation (Figure 2, top) and immunoblotting with organelle specific antibodies (Figure
2, bottom). You also collect small aliquots of the total cell extract and from each isolation step, and
with these immunoblotting using specific antibodies was carried out (Figure 3) and NDPase activity
was measured (Table 1).
Your task is to generate a short report to provide information on the efficiency of your Golgi
purification procedure.
In the short report, you will need to:
– State the purpose of this report.
– Briefly describe the key steps of Golgi isolation (in two or three sentences).
– Describe the efficiency of different steps of isolation procedure using all the datasets given.
In your text, you must refer to your figures. e.g. “the absence of a contaminating organelle
marker after FFE purification (Figure 1A)….”.
– Cite appropriate references to support your statements when necessary.