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Cat. No.: CIDC-50 (for 10-50T)

Cat. No.: CIDC-250 (for 50-250T)

Description

Chloroplast Isolation Kit by Differential Centrifugation is a reagent kit designed to rapidly extract intact and biologically active chloroplasts from fresh plant leaves within one hour using grinding, filtering, and differential centrifugation. The chloroplasts obtained with this kit have highly intact structures and can be used for biological function studies such as electron transport and phosphorylation in the photosynthetic electron chain, transmembrane transport and assembly of chloroplast proteins, as well as in vitro chloroplast protein synthesis and localization. Additionally, the chloroplasts can serve as starting material for the extraction of chloroplast proteins and the isolation and purification of chloroplast DNA and RNA.

Chloroplasts are double-membrane organelles unique to certain algae and higher plants that perform photosynthesis and are involved in various metabolic processes such as amino acid synthesis, chlorophyll synthesis, and lipid biosynthesis. In higher plants, chloroplasts are lens-shaped, with a diameter of approximately 5-10μm and a thickness of about 1-3μm. Typically, there are about 50-200 chloroplasts per mesophyll cell, accounting for roughly 40% of the cell’s cytoplasmic volume. Although the majority of chloroplasts are found in leaves, they are also present in stems, seeds, and immature fruits.

This kit uses differential centrifugation to isolate intact chloroplasts.

Components

• Chloroplast Extraction Buffer I
• Chloroplast Extraction Buffer II
• Chloroplast Preservation Solution
• Cell Strainers

Features

• This kit has a wide range of applications. It is broadly applicable to various types of fresh plants, including monocotyledons (such as Epipremnum aureum, maize, wheat) and dicotyledons (such as Arabidopsis thaliana, tobacco, spinach, soybean, and cabbage). 
• The kit is easy to use, requiring no special disruption equipment, and can quickly obtain high-purity, intact, biologically active chloroplasts within one hour.
• This kit provides a high yield of chloroplasts. Using this product, approximately 0.1-0.2 mg of intact chloroplasts can be obtained from each gram of leaves. For example, with Epipremnum aureum, approximately 0.15 mg of intact chloroplasts can be obtained from each gram of leaves using this product.
• Considering processing 0.2g or 1g of leaves per extraction, the small and medium packages of this kit can perform 10-50 and 50-250 extractions, respectively. The filters included in the kit are sufficient for only 10 and 50 extractions; for additional filters, it is recommended to purchase Cell Filters (70μm pore size, individually packaged, sterile).

Storage Conditions

Store at -20ºC, valid for one year. Store Cell Strainers at room temperature.

Precautions

• Acetone, ultrapure water or deionized water, scissors, centrifuge tubes, and disposable plastic pestles or glass homogenizers, mortars, and homogenizers are required. It is recommended to use Ultrapure Water (DNase/RNase-Free, Sterile).
• Chloroplasts are highly sensitive to temperature; therefore, all steps in the chloroplast isolation process must be performed on ice, and all solutions should be ice-cooled. Centrifugation should be carried out using a 4ºC centrifuge.
• During resuspension, if using standard pipette tips, cut off the tip to increase the diameter and gently pipette to avoid vigorous action that could rupture the chloroplasts.
• Throughout the experimental procedures, avoid vigorous shaking or pipetting. Handle gently and slowly to prevent chloroplast rupture.
• The chloroplast resuspension obtained is very prone to losing activity, so it is advisable to use it as soon as possible if performing activity assays.
• Thawed reagents must be completely thawed and mixed before use.
• If more cell filters are needed, it is recommended to purchase Cell Strainers (70μm pore size, individually packaged, sterile).
• This product is for scientific research use by professionals only. It is not intended for clinical diagnosis or treatment, nor for food or drug use, and should not be stored in residential areas.
• For your safety and health, please wear a lab coat and disposable gloves while handling.