10×ACK Lysis Buffer

10×ACK Lysis Buffer

Introduction

Red Blood Cell Lysis Buffer (RBC Lysis), also known as ACK Lysis Buffer, is a kind of solution used to lyse and remove enucleated red blood cells from blood or tissue samples of human or mouse.

This product is sterile. After treating with this buffer, the single cell suspensions can be used for subsequent primary culture, cell fusion, extraction of nucleic acid or protein, and various routine analysis and detection.

Instructions

10×ACK Lysis Buffer is concentrated, dilute with ddH₂O to 1× working solution before use.

For example: take 1 mL 10× ACK Lysis Buffer [E-CK-A105], add to 9 mL ddH₂O.The mixture is 1× ACK Lysis Working Buffer.

Experimental Procedure

For Mouse Spleen or Bone Marrow Samples

1. Centrifuge single cell suspension of the sample at 300 g for 5 min, discard the supernatant.
2. Add 2 mL of 1 ×ACK Lysis Working Buffer to the sample to resuspend the cells, and incubate at RT for 2~3 min (Incubation time should be adjusted accordingly to the actual situation).
3. Add 10 mL PBS to stop the lysis reaction.
4. Centrifuge at 300 g for 5 min, discard the supernatant.
5. Resuspend the cells with PBS or similar physiological buffer for subsequent experiments.

For Human Peripheral Blood Samples

1. Add 100 μL fresh blood into a centrifuge tube, add 2 mL of 1 ×ACK Lysis Working Buffer and mix thoroughly.
2. Put the centrifuge tube at 4°C for 10 min, until the fresh blood becomes transparent.
3. Centrifuge at 300 g for 5 min, discard the supernatant. Use a pipette to remove the residual supernatant carefully.
4. Add 2 mL PBS to the tube and repeat step 3.
5. Resuspend the cells with PBS or similar physiological buffer for subsequent experiments.

Storage

Store at 2~8°C. Avoid of freezing. Valid for 12 months.

Notes

1. This product is sterile. Please open and use it in laminar flow bench.
2. This reagent is for research use only.
3. For your safety and health, please wear the lab coat and disposable gloves before the experiments.