MonoZero™ Monocytes Blocking Reagent

Applications:

FC

Cat no : PF00020

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Validation Data Gallery

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  Human PBMCs were incubated with (bottom) and without (top) MonoZero™ Monocytes Blocking Reagent, then stained with FcZero-rAb™ Rabbit IgG Isotype Control Recombinant Antibody (FCA98136) conjugated to various Cyanine tandem fluorophores (from left to right: PE-Cyanine7, PE-Cyanine5.5, APC-Cyanine7, PerCP-Cyanine5.5).

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  MonoZero™ Monocytes Blocking Reagent, in staining with PE-Cyanine7 conjugated antibodies, shows significant reduction in background without affecting specific binding of antibody to the target protein, resulting in improved separation of positive and negative populations. Human PBMCs were incubated with (bottom) and without (top) MonoZero™ Monocytes Blocking Reagent, then stained with PY7-FcA65564 (CD73) (left), PY7-FcA65575 (CD20) (center), and PY7-FcA65621 (CD28) (right).

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  MonoZero™ Monocytes Blocking Reagent reduces non-specific background in mouse bone marrow without affecting specific binding of antibody to the target protein. Moues bone marrow cells were incubated with (bottom) and without (top) MonoZero™ Monocytes Blocking Reagent, then stained with PY7-FcA98136 (PE-Cyanine7 conjugated Rabbit IgG Isotype Control, left), or PY7-FcA65672 (PE-Cyanine7 conjugated Anti-Mouse CD11b, right).

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Product Information

In flow cytometry, both monocytes and macrophages exhibit non-specific binding to certain antibodies and fluorophores, in particular to anthocyanidin dyes such as Cyanine dyes and related tandem dyes such as PE-Cyanine 5.5, PE-Cyanine 5, PerCP-Cyanine 5.5, PE-Cyanine 7, and APC-Cyanine7. MonoZero™ Monocyte Blocking Reagent reduces non-specific binding of these cells populations to anthocyanidin dyes, increasing data quality and reproducibility while decreasing the complexity of data analysis.

MonoZero™ Monocyte Blocking Reagent is a non-antibody blocking reagent and does not affect the specific staining of monocytes and macrophages. For best results, use in conjunction with Fc Receptor Block or with FcZero-rAb™ Recombinant Antibodies for Flow Cytometry.

This product is for research use only.

Storage

Store sealed at 2-8℃ for up to 1 year and avoid repeated freeze thaw. Check that reagent is fully thawed and homogenous before use for best results.

Usage

This product is for use in flow cytometry experiments. The recommended titration is 1 uL per 100 uL cells, though the optimal titration may depend on the sample type and experimental design. Add MonoZero™ Monocyte Blocking Reagent to the cell suspension immediately before incubating with primary antibody. 

Protocol:

1. Suspend cells in Flow Cytometry Staining Buffer at a concentration of 1x10^6 cells/100 uL.

2. Add MonoZero™ Monocyte Blocking Reagent to the cell suspension at a ratio of 1:100.

3. Mix gently by pipetting, then stain with primary antibodies. Incubate for 30 minutes in the dark.

4. After incubation, wash twice with Flow Cytometry Staining Buffer, centrifuge and aspirate the supernatant.

5. Resuspend cells, then perform flow cytometry analysis.

Cited in Article as

PF00020, MonoZero™ Monocytes Blocking Reagent, Proteintech, IL, USA