BD, 570004, BD Pharmingen™ MonoBlock™ Leukocyte Staining Buffer

Reactivity: Human (QC Testing), Mouse (Tested in Development)
Application: Flow cytometry (Routinely Tested)
Storage Buffer: Aqueous buffered solution containing protein stabilizer and ≤0.09% sodium azide.
Regulatory Status: RUO
Preparation And Storage: Preparation And Storage Store undiluted at 4°C.
Recommended Assay Procedures: Recommended Assay Procedures For more information on MonoBlock™ Leukocyte Staining Buffer, please visit our website at: https://www.bdbiosciences.com/monoblock Protocols for Multicolor Immunofluorescent Surface Staining of Cells using BD Pharmingen™ MonoBlock™ Leukocyte Staining Buffer Protocol 1 for Staining Peripheral Blood Mononuclear Cells (PBMC) or Bulk Erythrocyte-lysed Whole Blood or Cell Samples Prepared from Lymphoid Tissues. a. First add 10 μL BD Pharmingen™ MonoBlock™ Leukocyte Staining Buffer to each staining tube or plate microwell followed by addition of fluorescent antibody or antibody cocktails and carefully vortex. b. Add 100 μL containing suspended human or mouse leukocytes (eg, at ~10^6 cells) to each tube or microwell and carefully vortex. Alternatively: Can first add 10 μL of BD Pharmingen™ MonoBlock™ Leukocyte Staining Buffer and 100 μL containing suspended leukocytes to each tube or microwell and carefully vortex. Then add fluorescent antibody or antibody cocktail to the mixture of cells and blocking buffer. c. Incubate the suspended cells (30 min) protected from light at room temperature (RT) for human cells and at 4°C for mouse cells. d. After incubation, wash the cells twice with BD Pharmingen™ Stain Buffer (FBS) [Cat. No. 554656]. e. Resuspend cells in 500 μL BD Pharmingen™ Stain Buffer (FBS) for flow cytometric analysis. Protocol 2 for Staining Human Whole Blood a. First add 10 μL BD Pharmingen™ MonoBlock™ Leukocyte Staining Buffer to each staining tube followed by addition of fluorescent antibody or antibody cocktails and carefully vortex. b. Add 100 μL of human whole blood. Alternatively: Can first add 10 μL of BD Pharmingen™ MonoBlock™ Leukocyte Staining Buffer and 100 μL containing suspended human whole blood to each tube or microwell and carefully vortex. Then add fluorescent antibody or antibody cocktail to the mixture of cells and blocking buffer. c. Incubate the suspended cells  (30 min) at room temperature (RT) protected from light. d. After incubation, lyse erythrocytes with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) or BD FACS™ Lysing Solution (Cat. No. 349202). e. Wash the cells twice with BD Pharmingen™ Stain Buffer (FBS) [Cat. No. 554656]. f. Resuspend cells in 500 μL BD Pharmingen™ Stain Buffer (FBS) for flow cytometric analysis. Protocol Notes: • For human leukocytes, further reduction in potential nonspecific staining caused by fluorescent antibody binding to cellular Fc receptors can be reduced by preincubating the cells with BD Pharmingen™ Human BD Fc Block™ (Cat. No. 564219/564220). • For mouse leukocytes, further reduction in potential nonspecific staining caused by fluorescent antibody binding to cellular Fc receptors can similarly be reduced by preincubating the cells with BD Pharmingen™ Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142]. • BD Pharmingen™ MonoBlock™ Leukocyte Staining Buffer is compatible with BD Horizon™ Brilliant Stain Buffer (Cat. No. 566349 563749) and BD Horizon™ Brilliant Stain Buffer plus (Cat. No. 566385 568264). Data not shown but available upon request.