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A chloroplast, delimited by a two-membrane envelope, is a type of plastid that serves as the site of

photosynthesis. The photosynthetic function is based on developing an operational extensive internal

membrane network, called the thylakoids, and on enzymatic processes in the chloroplast matrix, called the

stroma. Thylakoid membrane is structurally different from the chloroplast envelope, and their biogenesis

depends on biosynthetic and transporting activities specific to the chloroplast envelope.This kit is designed to

fractionate chloroplasts into three parts (thylakoid membrane, envelope membrane, and stroma) using a

specialized filter cartridge and differential centrifugation (without ultracentrifugation). Isolated thylakoids are in

native forms and can be used for functional studies and the studies of subcellular localization of proteins

using Western blotting/Mass spectrometry. The buffers in the kit are detergent and EDTA-free. The protocol

can be completed in about one hour.

Fig.1. Fractionation of chloroplasts from spinach leaf into three fractions: Thylakoid (T), Stroma (S), and

Envelope membranes (E).

Fig. 2. SDS-PAGE (Upper) and Western blot (Lower) of fractionated chloroplasts.   Lane 1. Total tissue

lysate, 2. Thylakoid, 3. Stroma and 4. Envelope membranes. Thylakoid marker antibody was purchased from

Agrisera (AS05084).