FceR1 alpha Monoclonal Antibody (AER-37 (CRA1)), NovaFluor™ Yellow 610, eBioscience™

Product Specific Information

Description
The AER-37 monoclonal antibody reacts with the Fc epsilon RI alpha subunit, an IgE-binding subunit lacking signal-transducing ability.

This product contains 1 vial of NovaFluor conjugate and 1 vial of CellBlox Plus Blocking Buffer.

Applications Tested
This AER-37 (CRA1) antibody has been pre-titrated and tested by flow cytometric analysis of peripheral blood cells. This can be used at 5 µL (0.2 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.

Master mixes
• Master mixes of NFs should be made at 2-8 °C and may be made up to 4 hours ahead of time.
• We do not recommend storing master mixes containing NovaFluor conjugates overnight or longer.

Whole Blood compatibility
• When utilizing whole blood (as opposed to density-gradient-purified PBMC), we recommend lysing red blood cells in bulk prior to staining with NovaFluor conjugates.
• See the Bulk Lysis of Human Whole Blood protocol here.
• Staining of whole blood with NovaFluor conjugates followed by lysis of red blood cells may result in higher-than-expected background staining.

Viability dye compatibility
• NovaFluor dyes are not compatible with DNA intercalating viability dyes.
• Do not use viability dyes such as propidium iodide, 7-actinomycin D (7-AAD) and DAPI. Invitrogen LIVE/DEAD Fixable Dead Cell stains are recommended for use with NovaFluor dyes.

CellBlox Plus Blocking Buffer
• This NovaFluor conjugate comes with CellBlox Plus Blocking Buffer (for 25 µg or 25 test products, Cat. No. C001T02F01; for 100 µg or 100 test, Cat. No. C001T03F01), essential for optimal staining.
• Use CellBlox Plus Blocking Buffer in all experiments with NovaFluor conjugates.
• Add 5 µL per sample to antibody cocktails/master mixes (regardless of how many Novafluor-conjugated antibodies are present) before combining with cells.
• CellBlox Plus Blocking Buffer is compatible with either Super Bright Complete Blocking Buffer or Brilliant Stain Buffer and can be used in antibody cocktails/master mixes with those reagents.
• For single-color controls, use 5 µL of CellBlox Plus Blocking Buffer per 100 µL of cell sample (10^3 to 10^8 cells).

NovaFluor conjugates are based on Phiton technology utilizing novel fluorophore-containing nucleic acid dye structures that allow for engineered fluorescent signatures with consideration for spillover and spread impacts. Learn more

Our internal testing shows that NovaFluor Yellow 610 non-specifically stains B cells in SJL mice. Non-specific staining has not been observed in BALB/c or C57BL/6 mice. Other strains have not been tested. See the Antibody Testing Data for an example of this strain-dependent difference.

Excitation: 551 nm; Emission: 614 nm; Laser: 561 nm (Yellow) Laser

Target Information

Fc epsilon RI alpha, also known as FceR1 alpha, is a subunit of the high-affinity receptor for IgE, primarily expressed on mast cells and basophils. Its expression is upregulated by the presence of IgE. Fc epsilon RI alpha forms a tetrameric complex with one beta and two gamma subunits, which are essential for signal transduction. The beta and gamma subunits contain immunoreceptor tyrosine-based activation motifs (ITAMs). The Fc epsilon RI complex plays a crucial role in triggering IgE-mediated allergic reactions. When allergen-bound IgE molecules bring together two or more high-affinity IgE receptors, it leads to the release of mediators such as histamine, which are responsible for allergy symptoms. This process couples allergens and mast cells to initiate inflammatory responses characteristic of allergic disorders like hay fever and asthma. The release of histamine and proteases also leads to the synthesis of prostaglandins and leukotrienes, which are potent effectors of the hypersensitivity response.