Human Fc Receptor Blocking Solution - sterile-filtered, BioReagent, 10 μL/test

Cat. No.: H1374078

GRADE & PURITY Sterile-filtered, BioReagent, 10 μL/test

Synonyms

Cyanine dye blocking solution | Blocking buffer for Flow | Blocking Reagent | Monocyte Blocker | Fc Receptor Blocker | FcR Blocking Reagent | Fc Receptor Blocking Solution

Storage

Store at 2-8°C,Protected from light

Shipped In

Wet ice

Application

Flow Cytometry

Overview

Non-specific monocyte binding remains a ubiquitous challenge in flow cytometry. Although the exact mechanism is undefined, literature implicates CD64 interactions with cyanine dyes. However, our data demonstrate that non-specific binding extends beyond cyanine-based fluorophores: certain non-cyanine fluorescent dyes also exhibit this phenomenon. Crucially, we observed selective non-specific staining on monocytes for all tested PE and APC tandem dyes. Human Fc Receptor Blocking Solution is a non-antibody blocking solution and has been formulated to block non-specific binding of PE-Cy7, PE-Cy5, PE-Cy5.5, PE-APC, APC-Cy7, and AF647 which is also a cyanine dye commonly appeared on monocytes and macrophages.

This buffer contains specialized human IgG containing 0.05% ProClin 300. It is not recommended to be used for staining human IgG. Handle as biohazard agent.

Human peripheral blood leukocytes were either not preincubated (Upper Plots) or were preincubated (Lower Plots) with Human Fc Receptor Blocking Solution as indicated. The cells were then selectively stained with either Mouse IgG1-kappa (PE-Cy5.5), anti-E8L (Ab216584), Mouse IgG1-kappa (PE-Cy7), anti-E8L (Ab216583), Mouse IgG2b-kappa (PE-Cy5), anti-E8L (Ab216565), Mouse IgG2c-kappa (PE-Cy5), anti-HEL (Ab212231) or Mouse IgG2c-kappa (PE-Cy7), ​anti-HEL (Ab212232). Two-dimensional pseudocolor plots of SSC-A vs Mouse IgG show that the Human Fc Receptor Blocking Solution reduces non-specific binding compared to staining without any blocking reagent. 

Human peripheral blood leukocytes were either not preincubated any blocking reagent (Top Plots) or were preincubated with competitor Fc Receptor Blocking reagent (Middle Plots) or were preincubated with Human Fc Receptor Blocking Solution (Bottom Plots) as indicated. The cells were then selectively stained with either Mouse IgG (APC-Cy7) (Ab213860), Mouse IgG2b-kappa (PE-Cy5), anti-E8L (Ab216565) or Mouse IgG2c-kappa (PE-Cy5), anti-HEL (Ab212231). Two-dimensional pseudocolor plots of SSC-A vs Mouse IgG show that the Human Fc Receptor Blocking Solution reduces non-specific binding compared to staining without any blocking reagent or with competitor Fc Receptor Blocking reagent. 

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Specifications

Synonyms

Cyanine dye blocking solution | Blocking buffer for Flow | Blocking Reagent | Monocyte Blocker | Fc Receptor Blocker | FcR Blocking Reagent | Fc Receptor Blocking Solution

Specifications & Purity

sterile-filtered, BioReagent, 10 μL/test

Stability And Storage

Store at 2-8°C for 12 months. When stored at 2°C to 8°C, the presence of serum in this product may cause aggregation of various proteins and lipoproteins (such as cold agglutinins, fibrinogen, vitronectin, etc.), forming visible precipitate or cloudiness.

Storage

Store at 2-8°C, Protected from light

Shipped In

Wet ice

This product requires cold chain shipping. Ground and other economy services are not available.

Grade

BioReagent, Sterile-filtered

Images

Flow Cytometry: Human Fc Receptor Blocking Solution (H1374078)

Human Fc Receptor Blocking Solution (H1374078) - Flow Cytometry
Human peripheral blood mononuclear cells were either untreated (black) or treated with Human Fc Receptor Blocking Solution (H1374078) (red) and stained with recombinant CD14 Antibody (AF405) (Ab214465) and CD3 Mouse mAb (PE-Cy5) (Ab189665) (left), CD3 Mouse mAb (PE-Cy7) (Ab189666) (middle), or CD3 Mouse mAb (AF647) (Ab189663).

Flow Cytometry: Human Fc Receptor Blocking Solution (H1374078)

Human Fc Receptor Blocking Solution (H1374078) - Flow Cytometry
Flow Cytometry analysis of no Human Fc Receptor Blocking Solution (left) or with Human Fc Receptor Blocking Solution (H1374078) (right) blocked human peripheral blood monocyte labelling CD11b (red) with CD11b Mouse mAb (PE-Cy5) (Ab191617) at 1.1 μg/mL for 1 hour at 4°C. Blue - Mouse IgG (PE-Cy5) (Ab190078). Black - Unlabelled control, cells without incubation with primary antibody.

Flow Cytometry: Human Fc Receptor Blocking Solution (H1374078)

Human Fc Receptor Blocking Solution (H1374078) - Flow Cytometry
Human peripheral blood leukocytes were either not preincubated (Left Plots) or were preincubated (Right Plots) with Human Fc Receptor Blocking Solution (H1374078) as indicated. The cells were then stained with Mouse IgG (PE-Cy5) (Ab190078). Two-dimensional pseudocolor plots of SSC-A vs Mouse IgG show that the Human Fc Receptor Blocking Solution reduces non-specific binding compared to staining without any blocking reagent.

Application

• Application: Dilution info
• Flow Cytometry:

  10 μL/test.; It is recommended to add 10 µL of Human Fc Receptor Blocking Solution per million cells in 100 µl staining volume, mix and incubate at 4 °C for 30 minutes prior to staining with antibody of inter