Before use, dilute the 10X red blood cell lysis buffer with sterile deionized water to a 1X working solution, and equilibrate the lysis buffer to room temperature prior to use.
For tissue cell samples:
1. Fresh tissue is digested with collagenase or trypsin, then dispersed into a cell suspension by an appropriate method. Centrifuge at 1500 rpm for 3 minutes and discard the supernatant.
2. Add 3–5 times the volume of cells in red blood cell lysis buffer, gently pipette to mix, and lyse at room temperature for 3 minutes.
3. Add an equal volume of buffer (such as PBS, pH 7.2–7.4), centrifuge at 4°C at 3000 rpm for 5 minutes, and discard the reddish supernatant.
4. If red blood cell lysis is incomplete, repeat steps 2 and 3 once. Typically, a very small amount of residual red blood cells will not affect subsequent experiments.
5. Wash 1–2 times with an appropriate buffer. Resuspend the pellet, centrifuge at 4°C at 1500 rpm for 2–3 minutes, and discard the supernatant.
6. After resuspending the cell pellet in a suitable solution, proceed with subsequent operations.
For blood samples:
1. Take fresh anticoagulated blood, add red blood cell lysis buffer at a volume 5–10 times that of the cells, gently pipette to mix, and lyse at room temperature for 3–5 minutes.
2. Add an equal volume of buffer (such as PBS, pH 7.2–7.4), centrifuge at 4°C at 3000 rpm for 5 minutes, and discard the reddish supernatant.
3. If incomplete red blood cell lysis is observed, repeat steps 2 and 3 once. Typically, a very small amount of residual red blood cells will not affect subsequent experiments.
4. Wash 1–2 times with an appropriate buffer. Resuspend the pellet, centrifuge at 4ºC at 1500 rpm for 2–3 minutes, and discard the supernatant.
5. After resuspending the cell pellet in the appropriate solution, you can proceed with subsequent operations.
Red Blood Cell Lysis Buffer (10X)
Red Blood Cell Lysis Buffer (10X)
Product Details
Product Details
Product Specification
| Conjugation | Unconjugated |
| Physical Appearance | Liquid |
| Stability & Storage | 12 months from date of receipt / reconstitution, 2 to 8 °C as supplied |
Background
This product, Red Blood Cell Lysis Buffer, is a solution used to lyse and remove anucleate red blood cells from fresh blood or tissue samples of humans, mice, rats, and other species. Through formula optimization, it efficiently lyses red blood cells while causing minimal damage to lymphocytes or other nucleated cells. Under normal conditions, this product can recover more than 80% of the total white blood cells in the sample, with cell viability remaining unaffected. The lysis buffer has been sterilized, so the isolated cell suspension can be directly used for subsequent applications such as primary cell culture, cell fusion, and flow cytometric sorting. It can also be directly employed for the extraction of nucleic acids or proteins, as well as various routine analyses and assays. The main component of this lysis buffer is ammonium chloride, containing less than 4% fixative.
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Contents & Protocol
