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Invitrogen
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Description
The monoclonal antibody 42D1 recognizes the CD85d protein also known as ILT4, LIR-2. 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.
This product contains 1 vial of NovaFluor conjugate and 1 vial of CellBlox Plus Blocking Buffer.
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.
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 (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: 637 nm; Emission: 713 nm; Laser: 633-640 nm (Red) Laser
CD85d, also known as LILRB2 or ILT4, is a member of the immunoglobulin-like transcript (ILT), leukocyte Ig-like receptor (LIR), and monocyte Ig-like receptor (MIR) family. It is a 110 kDa single transmembrane receptor with three Ig-like domains and a long cytoplasmic domain containing immunoreceptor tyrosine-based inhibitory motifs (ITIMs). CD85d is expressed on monocytes, macrophages, and dendritic cells. The receptor binds to non-classical class I molecules such as HLA-G, -B, and G1. Upon ligand binding, the tyrosine phosphatase SHP-1 interacts with the ITIMs, transducing a negative signal that inhibits immune response stimulation. This interaction is thought to contribute to tolerance during fetal implantation and plays a role in controlling inflammatory responses and cytotoxicity, helping to focus the immune response and limit autoreactivity. CD85d is also involved in organ allograft rejection, with the levels of ILT3 and ILT4 on dendritic cells suggesting its role in this process. The LILRB2 gene, located in a gene cluster at chromosomal region 19q13.4, encodes multiple transcript variants with different isoforms, highlighting its complexity and importance in immune regulation.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Watch the video to learn how to use the Invitrogen Flow Cytometry Panel Builder to build your next flow cytometry panel in 5 easy steps.
Protein Aliases: CD85 antigen-like family member D; CD85d; CD85d antigen; Ig-like transcript 4; ILT-4; Immunoglobulin-like transcript 4; leucocyte Ig-like receptor B2; Leukocyte immunoglobulin-like receptor subfamily B member 2; leukocyte immunoglobulin-like receptor, subfamily B (with TM and ITIM domains), member 2; LIR-2; LIR2CD85D; MIR-10; MIR10LILRA6; Monocyte/macrophage immunoglobulin-like receptor 10; myeloid inhibitory receptor 10
Gene Aliases: CD85D; ILT-4; ILT4; LILRB2; LIR-2; LIR2; MIR-10; MIR10
UniProt ID: (Human) Q8N423
Entrez Gene ID: (Human) 10288
If an Invitrogen™ antibody doesn't perform as described on our website or datasheet,we'll replace the product at no cost to you, or provide you with a credit for a future purchase.*
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