Red Blood Cell Lysis Buffer (10X), For Lysing Erythrocytes (RBC)

Boster’s RBC Lysis Buffer is supplied as a 10X solution contains ammonium chloride, which lyses red blood cells with minimal effects on leukocytes or other cells with nucleus.

Product Info Summary

SKU:AR1118
Size:100 mL (The kit contains sufficient lysis buffer for 200 cell pellet fractions containing 5x 106 cells each)
Physical State:Liquid
Application:The processed blood or tissue cell samples can be used for subsequent primary culture, cell fusion, nucleic acid or protein extraction, and various conventional analysis and detection.

Overview

Product Name RBC Lysis Buffer (10X)
SKU/Catalog Number AR1118
Pack Size 100 mL (The kit contains sufficient lysis buffer for 200 cell pellet fractions containing 5x 106 cells each)
Description Boster’s RBC Lysis Buffer is supplied as a 10X solution contains ammonium chloride, which lyses red blood cells with minimal effects on leukocytes or other cells with nucleus.
Storage & Expiration Upon receipt store at 4°C. RBC Lysis Buffer (10X) is stable for one year. Product is shipped on ice.
Equivalent Thermofisher (Product No. 00-4300)
ApplicationThe processed blood or tissue cell samples can be used for subsequent primary culture, cell fusion, nucleic acid or protein extraction, and various conventional analysis and detection.
*Our Boster Guarantee covers the use of this product in the above tested applications.
Cite This Product RBC Lysis Buffer (10X) (Boster Biological Technology, Pleasanton CA, USA, Catalog # AR1118)

Description

RBC (Red Blood Cell) Lysis Buffer has been designed, formulated, and tested to ensure optimal lysis of RBCs in single cell suspensions of blood or tissue samples from human or mouse. RBC Lysis Buffer is supplied as a 10X solution contains ammonium chloride, which lyses red blood cells with minimal effects on leukocytes or other cells with nucleus. Nucleated RBCs are not effectively lysed with ammonium chloride. The buffer should be diluted in deionized water prior to use. It is sterile filtered, and the processed blood or tissue cell samples can be used for subsequent primary culture, cell fusion, nucleic acid or protein extraction, and various conventional analysis and detection.

Procedure for Lysis of Tissue Cells

RBC Lysis Buffer (10X) should be diluted in deionized water prior to use.

1. Digest fresh tissue samples and prepare a single-cell suspension.

2. Centrifuge harvested cell suspension at 400-500xg for 5min at 4°C, then carefully remove and discard the supernatant.

3. Add 1X prepared RBC Lysis Buffer to the cell pellet at 3:1 to 5:1 (i.e. Add 3-5 mL 1X prepared RBC Lysis Buffer to 1mL cell pellet.) Vortex briefly. Incubate on ice for 4-5 min. Shake gently during incubation.

4. Centrifuge at 400-500xg for 5min at 4°C, then carefully remove and discard the red supernatant.

5. (Optional) Repeat Step 4 and Step 5 if further removal of red blood cells is needed. Small numbers of residual red blood cells do not interfere with subsequent assays.

6. Resuspend the cell pellet in at least 5 volume PBS, HBSS, normal saline or serum free medium for 1-2 times. Centrifuge at 400-500xg for 2-3 min at 4°C, then carefully remove and discard the supernatant.

7. Count the cells, adjust the density, and stain the cells.

Procedure for Lysis of Whole Blood

RBC Lysis Buffer (10X) should be diluted in deionized water prior to use.

1. Centrifuge harvested fresh anticoagulant blood at 400-500xg for 5min at 4°C, then carefully remove and discard the supernatant.

2. Add 1X prepared RBC Lysis Buffer to the cell pellet at 10:1 (i.e. Add 10 mL 1X prepared RBC Lysis Buffer to 1mL cell pellet.) Vortex briefly. Incubate on ice for 4-5 min. Shake gently during incubation.

3. Centrifuge at 400-500xg for 5min at 4°C, then carefully remove and discard the red supernatant.

4. (Optional) Repeat Step 2 and Step 3 if further removal of red blood cells is needed. Small numbers of residual red blood cells do not interfere with subsequent assays.

5. Resuspend the cell pellet in at least 5 volume PBS, HBSS, normal saline or serum free medium for 1-2 times. Centrifuge at 400-500xg for 2-3 min at 4°C, then carefully remove and discard the supernatant.

6. Count the cells, adjust the density, and stain the cells.

Notes:
For a small number of blood cells, Step 1 can be omitted. In Step 2, for mouse blood, it is sufficient to incubate on ice for 4-5 min. For human peripheral blood, it is better to extend the incubation time to 10 min, but no longer than 15 min. Shake gently during incubation. Step 4 to Step 6 remain the same.

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