MonoZero™ Human Fc Blocking Reagent

• 

  Human PBMCs were incubated with (right) and without (center) Human Fc Receptor Blocking Reagent, then incubated with PE-98136 (Rabbit IgG Isotype Control). Human Fc Receptor Blocking Reagent significantly reduces Fc-receptor mediated non-specific binding and improved signal-to-noise ratio.

Cat No. PF00032

Reactivity

Human

Application

FC

---

Product Information

FcγR is a type I transmembrane protein of the immunoglobulin superfamily. There are three subtypes, FcγRI (CD64), FcγRII (CD32) and FcγRIII (CD16), which are expressed on cells such as B cells, monocytes, macrophages, NK cells, and granulocytes. In immunology experiments such as cell sorting, immunofluorescence, immunohistochemistry, and flow cytometry, the Fc regions of primary and secondary antibodies can non-specifically bind to Fcγ receptors on the cell surface, causing high background and leading to significant errors in experimental results. Human Fc Receptor Blocking Reagent can be used to block the binding of Fc receptors to antibodies to achieve more accurate experimental results.

This product is a chimeric antibody with a porcine IgG1 constant region and should not affect the binding of primary and secondary antibodies in the experiment. This product also contains a human IgG Fc fragment. This product is not recommended for experiments that include anti-human IgG Fc antibodies. This product is not compatible with CD16/CD32/CD64 staining using antibodies whose epitope is at the binding interface.

Components

Buffer: PBS with 0.1% Proclin300, pH 7.2

Storage

Store at 2-8℃ for 1 year and avoid freezing. Before use, fully thaw and thoroughly mix the product for optimal performance. Avoid prolonged exposure to room temperature. After opening, the product should be stored sealed to prevent contamination.

Usage

This product is suitable for flow cytometry experiments. The recommended concentration is 1.0 uL Fc Receptor Blocking Reagent per 100 uL of cell suspension, though concentration may be adjusted depending on sample and experimental protocol. When using, first mix the cell suspension with Fc Blocking Reagent in the recommended ratio, then immediately add the antibody for incubation.

Protocol:

1. Suspend cells at approximately 1x10^7 cells/mL.

2. Add 1.0 uL Human Fc Receptor Blocking Reagent per 100 uL cells. Gently mix and incubate at room temperature for 10 minutes.

3. Add primary antibodies and incubate for 30 minutes.

4. Wash twice with flow cytometry staining buffer (PF00018). Centrifuge and discard the supernatant.

5. Resuspend the cells and proceed with flow cytometry analysis.

Note

This product is recommended for use only on human cells.

Cited in Article as

PF00032, MonoZero™ Human Fc Blocking Reagent, Proteintech, IL, USA