Storage and Handling
Store at room temperature and protected from sunlight.
Stable for 12 months from the date of manufacture.
Leinco Technologies' RBC Lyse Buffer, “Ready to Use” has been designed to gently lyse erythrocytes while maintaining exquisite populations of leukocytes. Comparable to the ACK (Ammonium-Chloride-Potassium) Lysing Buffer this product can be used for the lysis of red blood cells in samples containing white blood cells, such as EDTA-treated whole blood, buffy coats, and bone marrow. RBC Lysing Buffer, “Ready to Use” is manufactured at a cGMP-compliant facility located in Saint Louis, Missouri. The facility is registered and is certified to ISO 9001.2008 standards.
Reagents: 1X RBC Lysing Buffer
Working Dilutions: This product is provided at a 1X working concentration and is Ready to Use.
Directions for Use
1. Receipt of the timed specimen collected in sodium heparin or EDTA and an automated leukocyte count and differential can be performed.
2. Whole Blood is to be pipetted (1-3 ml) into a 15 ml conical centrifuge tube, along with approximately 12ml phosphate buffered saline (PBS) containing 10 units/ml heparin and centrifuged for 5 min at 400g. The clear supernatant should be aspirated to within 500 μl of the cell pellet.
3. The cells are to be washed a second time with PBS (without heparin) and resuspended in 1 ml of PBS containing 100 µg/ml normal mouse IgG (Leinco Part Number N229) to block Fc antibody binding.
4. Cells are to be incubated with IgG block on ice for 10-minute and then 100μl of cells are added to tubes containing cocktails of fluorochrome labeled mAbs or single directly conjugated antibodies.
NOTE: All mAbs were pretitered & used at saturating concentrations.
5. The sample tubes are mixed, and returned to the ice bath for 30 min.
6. After the incubation, 1 - 3.5 ml of lysing solution is added to each tube, the tubes inverted three times and held at room temperature for 5-6 minutes to promote lysis of erythrocytes.
7. FIXING OPTION: The tubes are centrifuged for 5 minutes at 400g, and washed once with PBS before resuspending in 350μl of 0.5% methanol free formaldehyde.
NOTE: Stained samples are stored in the dark at 2 -8oC for no longer than 3 days before analysis.
1.) Blaha, Michael & DuBose, David (2002). A HUMAN WHOLE BLOOD MODEL FOR SCREENING POTENTIAL VESICANT ANTAGONISTS. Natick, MA: US Army Research Institute of Environmental Medicine. Report Link
Products are for research use only. Not for use in diagnostic or therapeutic procedures.