CD5 Monoclonal Antibody (53-7.3), NovaFluor™ Blue 800

Product Specific Information

Description
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.

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

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.

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

Excitation: 493 nm; Emission: 806 nm; Laser: 488 nm (Blue) Laser

Target Information

CD5 is a 67 kDa human T-lymphocyte single-chain transmembrane glycoprotein. CD5 is present on all mature T-lymphocytes, on most of thymocytes and on many T-cell leukemias and lymphomas. CD5 also reacts with a subpopulation of activated B-cells and may act as a receptor in regulating T-cell proliferation. CD5 is found on 95% of thymocytes and 72% of peripheral blood lymphocytes. In lymph nodes, the main reactivity is observed in T cell areas. CD5 is expressed by many T cell leukemia, lymphomas, and activated T cells. Diseases associated with CD5 dysfunction include thymus cancer and Richter's Syndrome.