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Principle and Usage of Red Blood Cell Lysis Buffer
Red cell lysis buffer is a simple and convenient method for removing red blood cells: the buffer lysates the erythrocytes while leaving nucleated cells intact, thereby efficiently eliminating red cells. This gentle lysis approach is particularly suited for applications such as the isolation and purification of enzymatically digested tissue cells, the separation and purification of lymphocytes, and the removal of red blood cells in experiments involving the extraction of proteins and nucleic acids from tissue samples.
Principle Explanation
By exploiting the differences in ion concentrations between the inside and outside of the cell, an osmotic pressure gradient is established, causing water from the external environment to diffuse into the cell, thereby swelling the red blood cells and inducing lysis.
Instructions for Use
1. Lysis of red blood cells
(1) Add 3 times the volume of red cell lysis buffer to 1 mL of fresh whole blood to be lysed (e.g., add 3 mL of red cell lysis buffer to 1 mL of fresh whole blood).
Gently vortex or invert to mix. [Note]: If processing fresh tissue cells, they must first be digested with trypsin or other enzymes to obtain a single-cell suspension, followed by centrifugation to pellet the cells.
Collect the cells and then proceed with subsequent steps.
(2) Incubate on ice for 15 min, gently vortexing to mix twice during this period. [Note]: After red blood cell lysis, the solution should be clear and transparent.
2. Collect white blood cells
(1) At 4°C, centrifuge at 450×g for 10 min to pellet the leukocytes, then carefully aspirate and discard the supernatant.
(2) Add twice the volume of red blood cell lysis buffer to the above pellet, and gently vortex to fully resuspend the leukocytes (e.g., if the initial blood volume is 1 mL, add…).
Add 2 mL of red blood cell lysis buffer.
(3) At 4°C, centrifuge at 450×g for 10 min to pellet the leukocytes, then carefully and completely aspirate the supernatant.
(4) Resuspend the cells for subsequent experiments. [Note]: If RNA is to be extracted, it is best to use RNase-free solutions starting from this step.
Red blood cell lysis solution product
[Name] Li Ji Bio
[Name] Red Blood Cell Lysis Buffer
[Item Number] AP01L106
[Specification] 100 mL
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Email:ztt0005@obiosh.com
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