CD23 Monoclonal Antibody (B3B4), NovaFluor™ Yellow 610, eBioscience™

Product Specific Information

Description
The B3B4 monoclonal antibody reacts with mouse CD23, a 45 kDa type II transmembrane glycoprotein. CD23 is expressed on resting conventional B cells, and its expression is modulated upon B-cell activation. B-1 cell lineage (CD5+ B cells) does not express CD23. Soluble forms of the antigen have been reported to be biologically active. CD23 is a low affinity receptor for IgE and is thought to play a role in the regulation of IgE response and B-cell activation. CD21 is thought to bind to CD23.

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

Applications Tested
This eBio4KB128 antibody has been tested by flow cytometric analysis of mouse splenocytes. This may be used at less than or equal to 0.4 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. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

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

CD23 is a 45 kDa glycoprotein that serves as a low-affinity receptor for IgE, playing a crucial role in regulating IgE responses and B cell activation. It is expressed on mature B cells, mantle zone B cells, follicular dendritic cells, and at lower levels on T cells, NK cells, Langerhans cells, and platelets. CD23 expression is upregulated upon B cell activation, and its soluble forms are biologically active, acting as potent mitogenic factors. CD23 is strongly expressed on Epstein-Barr virus (EBV)-transformed B lymphoblasts and is present on a subpopulation of freshly isolated peripheral blood and tonsil B cells. It is also detected in neoplastic cells from cases of B cell chronic lymphocytic leukemia and some centroblastic/centrocytic lymphomas. Functionally, CD23 is involved in B cell growth, differentiation, and IgE production. It interacts with CD21 and the alpha subunits of CD11b and CD11c, further influencing immune responses. Diseases associated with CD23 dysfunction include chronic conjunctivitis and chronic lymphocytic leukemia, highlighting its importance in both normal immune function and disease states. This makes CD23 a valuable target for antibody customers interested in B-cell-related research and diagnostics.