Toll-Free Customer Service (855) 848-6626 Hours 6AM-5PM PST

FREE Shipping on orders over $300*

Your valued source for Immunology Tools

The Dragonfly Community

RBC Lysing Protocol

Posted by Michaela Frister on Aug 20, 2014 2:18:43 PM

RBC Lysis Buffer (10X) Cat. No. TNB-­4300-­L100

Other Materials Required

  • Flow Cytometry Staining Buffer (Stain Buffer) (1X PBS with 2% FBS, 0.09% Na-­Azide)

Note: This protocol applies to human whole blood preparations. For use with other tissue samples, it is recommended that the investigator optimize conditions to obtain best results. RBC Lysis Buffer (10X) has been tested using blood collected with either heparin or EDTA as the anti­coagulant and found to perform equivalently.

  1. Prepare 1X RBC Lysing Buffer by adding 1 part RBC Lysing Buffer (10X) with 9 parts room temperature distilled water.
  2. Aliquot a sample of whole blood, typically 50­-100 µL, to tube.
  3. Add fluorochrome­-labeled antibodies for staining directly to sample and mix thoroughly.
  4. Incubate for 30 minutes at room temperature and protected from light.
  5. Add 2 mL of room temperature 1X RBC Lysing Buffer (prepared in step 1) and pulse vortex (<5 seconds).
  6. Incubate for 10-­15 minutes at room temperature and protected from light.
  7. Centrifuge cells at 500 x g for 5 minutes at room temperature.
  8. Carefully decant supernatant and wash cells once with 1­-2 mL Stain Buffer.
  9. Centrifuge cells as in Step 7 and resuspend at the appropriate volume for analysis.

Download PDF here!

Topics: lysing, protocol, RBC, support

The Dragonfly Effect!

Be quick, effective, and powerful in your research

Tonbo Biosciences brings the tools to help take your research to the next level. Check back frequently to learn about the latest Tonbo offerings, tips, tricks, and events. Visit the Tonbo website to find the right tools for your immunology workflow. Sign up for our email alerts to keep updated on the latest from Tonbo.

  • Broad portfolio of research tools for the immunology workflow
  • Validated for consistent performance
  • Antibodies, Recombinants, PCR Reagents
  • Area of Focus: Immuno-Oncology, Innate, Adaptive, Allergy

Subscribe to Email Updates

Recent Posts