BioReagent BioReagent for sensitive chromatographic and analytical workflows requiring minimal baseline interference.
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Cited in 0 peer-reviewed publications across chromatography, organic synthesis, and cross-coupling reactions.
Chloroplasts are photosynthetic organelles. In photosynthesis research, extracted chloroplasts are often required for downstream studies. Chloroplast isolation provides an effective approach to obtain intact chloroplasts from plant leaves. The method includes mechanical disruption of cell walls and cell membranes, filtration for removal of leaf tissue and collection of cell debris, isolation of chloroplasts by centrifugation, and separation of intact chloroplasts from broken chloroplasts using layering solution or density‑gradient methods.
The Chloroplast Extraction Kit adopts a simple, well‑established and easy‑to‑perform experimental protocol to save experimental time. The isolated chloroplasts are suitable for studies on carbon assimilation, electron flow and phosphorylation, metabolic transport, protein targeting and other processes. Its extraction principle is as follows: leaf homogenate obtained by grinding leaves is subjected to filtration and centrifugation to harvest chloroplasts, whose integrity is then assayed in isotonic buffer. This kit is applicable for isolating chloroplasts from plant leaves (such as spinach, pea, lettuce, cabbage, chard and tobacco) for structural and functional research.
This kit is for research use only. It is not intended for clinical diagnosis or any other application
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Materials to be Prepared by User
1. Distilled water
2. Electronic balance, centrifuge tubes or test tubes, mortar or homogenizer, gauze or filter paper, refrigerated centrifuge, microscope
Operating Procedures (For Reference Only)
1. Select healthy plant leaves such as spinach preferably grown under several consecutive sunny days. Wash the leaves and remove the midribs, then pre‑cool them in a refrigerator at 0‑4 ℃.
2. Transfer 5 mL of Chloroplast Lysis Buffer into a mortar or homogenizer, and pre‑cool in refrigerator to near 0 ℃.
3. Weigh 2 g pre‑cooled sample, cut into pieces and place into the mortar. Grind rapidly by hand for 0.5‑1 min. (Note: Do not grind too hard or excessively fine; it is sufficient to crush leaves into small pieces.) Filter through 2‑3 layers of gauze or filter paper.
4. Transfer the filtrate into a pre‑cooled centrifuge tube, centrifuge at 300 g for 2 min at 4 ℃, discard the pellet.
5. Transfer the supernatant to another pre‑cooled centrifuge tube, centrifuge at 1000 g for 2 min at 4 ℃.
6. Discard the supernatant; the pellet is chloroplasts. Add 200‑400 μL pre‑cooled Chloroplast Suspension Solution to resuspend the pellet. Store at 4 ℃ for subsequent use.Observe chloroplast morphology under a microscope at 400‑600‑fold magnification.
Notes
1. Use healthy plant leaves preferably grown under several consecutive sunny days as samples for isolation.
2. Perform all operations at approximately 4 ℃ as much as possible.
3. Use the reagents promptly after opening to avoid compromised experimental performance.
4. For your safety and health, wear a lab coat and disposable gloves during operation.
Shelf‑life
Valid for 12 months. Ship under cold conditions; store at 4 ℃.
| Component No. | Component Name | Appearance | 50T | Storage |
| C1527488A | Chloroplast Lysis Buffer | Liquid | 250ml | 4℃ |
| C1527488B | Chloroplast suspension | Liquid | 20ml | 4℃. Store in the dark. |
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