Cell Lysates
Cell lysates are solutions containing the contents of cells that have been broken open, releasing their intracellular components. These lysates are commonly used in biological research to study the proteins, nucleic acids, and other molecules within a cell.
Cell lysates are commonly used tools in many areas of biological research, including molecular biology, cell biology, and biochemistry. They can be used to study protein expression levels, enzyme activity, pathway signaling, cancer diagnostics, and drug screening. By using cell lysates in western blot and ELISA experiments, researchers are able to gain valuable insights into the inner workings of cells and further our understanding of biological processes.
With a large number of cell products in stock, Creative Biolabs is passionate about providing high-quality, convenient, and cost-effective cell lysates prepared from primary cells, cell lines, and iPSC-derived cells. Please contact us to get cell lysate products of your interest.
Procedure
We usually use the following optimized process to obtain cell lysates.
1. Harvesting cells
Suspension cells: Collected directly by high-speed centrifugation at low temperature and washed with PBS to remove any remaining media or serum before further processing.
Adherent cells: Collected by scraping off to avoid potential protein loss.
2. Adding lysis buffer and ultrasonication
Lysis buffer: Modified RIPA Lysis Buffer is a gentle and effective solution for isolating cytoplasmic and nuclear proteins from mammalian cells. Free of SDS, this optimized buffer is ideal for proteins sensitive to harsh denaturing conditions, making it suitable for a wide range of applications such as ELISA-based assays.
Ultrasonication: By subjecting the sample to high-frequency sound waves, the cell membrane is disrupted, releasing the required biomolecules.
3. Lysate concentration determination
UV absorbance: UV absorbance is a common technique used to measure protein concentration in cell lysates. Protein molecules absorb light at a wavelength of 280nm, allowing for the quantification of protein concentration in cell lysates.
4. Lysate storage
Lysate is diluted and stored in Storage Buffer.
Applications
Then purified cell lysates can be used for a range of experiments:
- Immunodiagnostic tests for rapid and convenient on-site testing.
- Understanding the cellular heterogeneity.
- Down streaming processes, such as western blot and ELISA experiments.
Reference
- Shehadul Islam, Mohammed et al. "A Review on Macroscale and Microscale Cell Lysis Methods." Micromachines (Basel). 2017, 8(3):83.
Target
293T Cell Transient Overexpression
- Species:
- Human
- Applications:
- WB
- Cell Types:
- Other Cells
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