The 53-7.3 monoclonal antibody reacts with mouse CD5, a 67 kDa protein expressed by a majority of thymocytes, mature T cells and a subset of B cells. The expression of CD5 by a small subset of B cells characterizes a developmentally and functionally distinct lineage of B cells called B-1 cells. CD5 is a counter-receptor for CD72 and plays a role in the T-B cell interaction.

Applications Reported: The 53-7.3 antibody has been reported for use in flow cytometric analysis.

Applications Tested: The 53-7.3 antibody has been tested by flow cytometric analysis of mouse splenocytes. This can be used at less than or equal to 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. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest.

Our internal testing shows that NovaFluor Yellow 755 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.

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.

Each NovaFluor conjugate or kit is shipped with CellBlox Blocking Buffer. Use this buffer whenever staining with NovaFluor conjugates, including single-color compensation controls using cells. Whenever possible, we recommend adding CellBlox Blocking Buffer to antibody cocktails/master mixes prior to combining with cells. Add 5 µL per sample (regardless of the number of NovaFluors in your panel) to use the antibody cocktail as intended. For single-color controls, use 5 µL of CellBlox Blocking Buffer per 100µL of cell sample containing 10^3 to 10^8 cells.

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

Excitation: 552 nm; Emission: 755 nm; Laser: 561 nm (Yellow) Laser