The monoclonal antibody 42D1 recognizes the CD85d protein also known as ILT4, LIR-2. It is a member of the ILT (immunoglobulin-like transcript)/ LIR (leukocyte Ig-like receptor), MIR (monocytes Ig-like receptor) family. CD85d is a 110 kDa single transmembrane receptor with 3 Ig-like domains and a long cytoplasmic domain containing ITIMs (immunoreceptor tyrosine-based inhibitory motifs). Expression is confined to monocytes, macrophages and dendritic cells. The ligand for CD85d is non-classical class I HLA-G, -B and G1. Upon ligand binding, tyrosine phosphatase SHP-1 interacts with the ITIM. Interaction of HLA-G with ILT4/CD85d is thought to contribute to tolerance during fetal implantation. Additionally, it is the levels of ILT3 and ILT4 on DCs that suggests a role in organ allograft rejection. Crosslinking of the receptor with the monoclonal antibody 42D1 inhibits calcium flux as well as enhancing HLA-G tetramer staining and restoring the stimulating activity of tolerogenic DC.

Applications Reported: This 42D1 antibody has been reported for use in flow cytometric analysis.

Applications Tested: This 42D1 antibody has been pre-titrated and tested by flow cytometric analysis of normal human peripheral blood cells. This can be used at 5 µL (1 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.

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: 639 nm; Emission: 710 nm; Laser: 633-640 nm (Red) Laser