CD4 Monoclonal Antibody (RM4-5), NovaFluor™ Yellow 660, eBioscience™, Invitrogen™

Description

• CD4 Monoclonal antibody specifically detects CD4 in Mouse samples
• It is validated for Flow Cytometry Description: The RM4-5 monoclonal antibody reacts with the mouse CD4 molecule, a 55 kDa cell surface receptor expressed by a majority of thymocytes, subpopulation of mature T cells and dendritic cells
• CD4 binds to MHC class II on the surface of antigen presenting cells and plays an important role both in T cell development and in optimal functioning of mature T cells
• In T cells, CD4 associates with protein tyrosine kinase p56lck through its cytoplasmic tail
• Binding of RM4-5 is blocked by GK1.5
• Applications Reported: The RM4-5 antibody has been reported for use in flow cytometric analysis
• Applications Tested: The RM4-5 antibody has been tested by flow cytometric analysis of mouse thymocytes and splenocytes
• This can be used at less than or equal to 0.0125 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
• Each product contains 1 vial of NovaFluor conjugate and 1 vial of CellBlox Plus Blocking Buffer
• 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
• This NovaFluor conjugate has been updated to ship with CellBlox Plus Blocking Buffer (Cat
• No
• (<a href='https://www.thermofisher.com/order/catalog/product/C001T06F01'>C001T06F01</a>))
• This buffer contains formulation improvements over CellBlox
• CellBlox Plus Blocking Buffer is required for optimal staining with NovaFluor conjugates and should be used in all experiments where NovaFluor conjugates are used
• Whenever possible, we recommend adding CellBlox Plus 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 100 μL 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
• <a href = https://www.thermofisher.com/us/en/home/life-science/cell-analysis/flow-cytometry/novafluor-dyes.html>Learn more </a> Excitation: 552 nm; Emission: 663 nm; Laser: 561 nm (Yellow) Laser Our internal testing shows that NovaFluor Yellow 660 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.