Chloroplast Isolation Kit
SIGMA/CPISO - isolation of intact chloroplasts from leaves
Synonym: Chloroplast Purification Kit; Isolation Kit for Chloroplast; Kit for Chloroplast Isolation
Product Type: Chemical
Ferricyanide photoreduction – a measure of intact chloroplasts This assay is based upon the inability of the ferricyanide to cross the chloroplast envelope and to react with the electron transport system in the thylakoid membranes. Ferricyanide reduction, as indicated by the decrease in the
absorbancy at 410 nm, occurs only when ruptured chloroplasts are present in the
preparation.
The degree of integrity of the chloroplast preparation is assessed by comparing the
rate of ferricyanide reduction upon illumination before and after osmotic shock of
the chloroplasts.
Analysis of the results,presented here,indicates that 88% of the chloroplasts in
the spinach chloroplast preparation are intact.
Chloroplasts (equivalent to 25 mg/ml chlorophyll) prepared using CP-ISO were
illuminated in the presence of 1.5 mM ferricyanide. The reduction of ferricyanide
was measured spectrophotometrically (410 nm).
The graph demonsrates bars representing the slopes change in absorbancy of the differernt samples during 6 minutes.
absorbancy at 410 nm, occurs only when ruptured chloroplasts are present in the
preparation.
The degree of integrity of the chloroplast preparation is assessed by comparing the
rate of ferricyanide reduction upon illumination before and after osmotic shock of
the chloroplasts.
Analysis of the results,presented here,indicates that 88% of the chloroplasts in
the spinach chloroplast preparation are intact.
Chloroplasts (equivalent to 25 mg/ml chlorophyll) prepared using CP-ISO were
illuminated in the presence of 1.5 mM ferricyanide. The reduction of ferricyanide
was measured spectrophotometrically (410 nm).
The graph demonsrates bars representing the slopes change in absorbancy of the differernt samples during 6 minutes.
| Quality Level | 200  |
| shipped in | wet ice |
| storage temp. | 2-8°C |
| technique(s) | centrifugation: suitable |
| Application: | Chloroplast Isolation Kit may be used in chloroplast organelle research to isolate intact chloroplasts from plant leaves for structural and functional studies. |
| Application: | The Chloroplast Isolation Kit has been tested for use with spinach, pea, lettuce, cabbage, mangold, and tobacco. |
| Features and Benefits: | • Specifically formulated buffer - Saves time using easy to follow simple and proven protocols • Isolates intact chloroplasts - Useful for studies of such processes as carbon assimilation, electron flow and phosphorylation, metabolic transport or protein targeting • Total organelle protein complement - Useful for functional studies, metabolic transport, proteomic profiling, and genomic applications • Convenient format - Saves time and reduces waste |
| General description: | The Chloroplast Isolation Kit provides an efficient procedure for isolating intact chloroplasts from plant leaves. The procedure involves mechanical cell wall and cell membrane breakage, removal of cell debris and leaf tissue by filtration, collection of total cell chloroplast fraction by centrifugation, and separation of intact chloroplasts from broken chloroplasts using a Percoll® layer or gradient. |
| RIDADR | NONH for all modes of transport |
| Storage Temp. | 2-8°C |
| UNSPSC | 12352200 |
| Components | Chloroplast Isolation Buffer 5x (CIB) 500 mL; Bovine Serum Albumin, heat shock fraction, pH 5.2, ≥96% 3 g; Chloroplast Isolation Buffer 5x (CIB) 4 ea; Percoll®, pH 8.5-9.5 (25 °C), suitable for cell culture 100 mL |