Secretome
Secretome

Basic concepts

Pellet Chlorella cells, ppt secreted proteins from dilute solution

Run SDS gels and observe proteins

Mass spectrometry of proteins to determine what they are

Protocols

TCA ppt with a carrier is good (best?) way to recover dilute proteins

DOC and sodium lauryl sarcosinate (0.01-0.1%) are best carriers - see ref

Add DOC or SLS, bring to 10% in TCA (from 100% w/v solution)

Incubate on ice for an hour

Low speed centrifugation

Wash pellet with THF to remove detergent

Ref - Chevallet, et al - Proteomics 7, 1757-1770 (2007) - available online - see excerpt

Old pages
Feb 21

Feb 25 - info on running gels

TCA - potential carriers

Example of gel of proteins isolated by phenol extraction from dilute samples

Note that Coomassie blue seems better than dansyl chloride fluorescence. Carrier protein (ovalbumin) isn't helpful, at least with these samples

No protein bands that are obviously unique to lysin

Activities of above samples

Note that glycosidase activites and lysin activities are in different fractions