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Cat. No.: NCPE-50 (for 50T)

Cat. No.: NCPE-100 (for 100T)

Description

When studying cells, it is often necessary to investigate different components of the cells, with the most commonly studied being the cell nucleus and cytoplasm. Separating nuclear and cytoplasmic proteins not only allows for the study of protein localization within the cell but also provides nuclear proteins that can be used for research on transcriptional regulation, such as EMSA (also known as gel shift) and footprinting.

The Nuclear and Cytoplasmic Protein Extraction Kit provides a relatively simple and convenient method for extracting nuclear and cytoplasmic proteins from cultured cells or fresh tissue. The separation of nuclear and cytoplasmic proteins from cultured cells can be completed in approximately 90 minutes. The extracted proteins can be used for subsequent procedures such as Western blotting, EMSA, footprinting, reporter gene assays, and enzyme activity assays.

This kit utilizes the Cell Cytoplasmic Protein Extraction Reagents A and B to induce cell swelling under low osmotic conditions, followed by disruption of the cell membrane to release cytoplasmic proteins. The cell nuclei are then obtained by centrifugation, and the nuclear proteins are extracted using a high-salt nuclear protein extraction reagent.

For cell samples, the kit can extract proteins from up to 100 samples if the quantity of cells per sample does not exceed two million. For tissue samples, the kit can extract proteins from up to 100 samples if the weight of each sample does not exceed 30 milligrams. If each sample weighs approximately 30-60 milligrams, the kit can extract proteins from up to 75 samples.

Precautions

• Prepare PMSF separately. PMSF must be added to the extraction reagent within 2-3 minutes before adding it to the sample to avoid rapid inactivation in the aqueous solution.
• All steps of protein extraction should be performed on ice or at 4°C.
• This kit is only suitable for fresh tissue samples and is not effective for frozen tissue samples.
• The protein concentration of the extracted nuclear and cytoplasmic proteins can be directly measured using the BCA protein assay kit. However, the Bradford method is not suitable for protein concentration determination.
• This product is intended for scientific research purposes by professionals only. It should not be used for clinical diagnosis or treatment, food or drug purposes, and should not be stored in a regular household.
• For your safety and health, please wear appropriate laboratory attire and disposable gloves during operation.

Storage

Store at -20°C, valid for one year.

Related: Membrane and Cytosol Protein Extraction Kit

Only for research and not intended for treatment of humans or animals

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