Invitrogen

CD85j (ILT2) Monoclonal Antibody (HP-F1), NovaFluor™ Yellow 610, eBioscience™

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Datasheet
Protocols
Questions & Answers
Datasheet
Protocols
Questions & Answers

Product Details

H043T03Y03-A

Applications
Tested Dilution
Publications

Flow Cytometry (Flow)

5 µL (0.4 µg)/test
-
Product Specifications

Species Reactivity

Human

Host/Isotype

Mouse / IgG1, kappa

Class

Monoclonal

Type

Antibody

Clone

HP-F1

Conjugate

NovaFluor™ Yellow 610 NovaFluor™ Yellow 610 NovaFluor™ Yellow 610

Excitation/Emission Max

551/614 nm View spectra spectra

Form

Liquid

Concentration

5 µL/Test

Purification

Affinity chromatography

Storage buffer

PBS, pH 7.2

Contains

0.09% sodium azide

Storage conditions

4°C, store in dark, DO NOT FREEZE!

RRID

AB_3098536 (AB_2925933)

Product Specific Information

Description
The monoclonal antibody HP-F1 recognizes CD85j, also known as ILT2, LILRB1, and LIR1. The monoclonal antibody HP-F1 has been shown to reduce the amount of CD16- dependent cytolytic activity of functional NK cells.

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

Applications Tested
This HP-F1 antibody has been pre-diluted and tested by flow cytometric analysis of normal human peripheral blood cells. This may be used at 5 µL (0.25 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

Our internal testing shows that NovaFluor Yellow 610 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.

Excitation: 551 nm; Emission: 614 nm; Laser: 561 nm (Yellow) Laser

Target Information

CD85j, also known as a member of the leukocyte immunoglobulin-like receptor (LIR) family, is a single transmembrane glycoprotein with a long cytoplasmic domain containing four immunoreceptor tyrosine-based inhibitory motifs (ITIMs). These motifs signal through interactions with SHP-1, transducing negative signals that inhibit immune response stimulation. CD85j is expressed on myeloid cells such as monocytes and dendritic cells, as well as on subsets of lymphoid cells including NK, T, and B cells. The expression of CD85j is correlated with leukemias such as acute lymphoblastic leukemia (ALL) and chronic lymphocytic leukemia (CLL). On CD8+ cells, CD85j expression is associated with effector cell function and plays a significant role in viral infections, including HIV, Epstein-Barr virus, and cytomegalovirus (CMV). CD85j binds to MHC class I molecules, including HLA-G, A, F, B27, E, and F, on antigen-presenting cells. The receptor helps control inflammatory responses and cytotoxicity, focusing the immune response and limiting autoreactivity. CD85j belongs to the subfamily B class of LIR receptors, which contain two or four extracellular immunoglobulin domains, a transmembrane domain, and multiple transcript variants encoding different isoforms have been identified for this gene. Its gene is located in a cluster at chromosomal region 19q13.4.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

How to use the Panel Builder

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.

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Bioinformatics

Protein Aliases: CD85 antigen-like family member J; CD85j; Ig-like transcript 2; ILT-2; Immunoglobulin-like transcript 2; leucocyte Ig-like receptor B1; Leukocyte immunoglobulin-like receptor subfamily B member 1; leukocyte immunoglobulin-like receptor subfamily B member 1 soluble isoform; leukocyte immunoglobulin-like receptor, subfamily B (with TM and ITIM domains), member 1; LIR-1; MIR-7; Monocyte/macrophage immunoglobulin-like receptor 7; myeloid inhibitory receptor 7

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Gene Aliases: CD85J; ILT-2; ILT2; LILRB1; LIR-1; LIR1; MIR-7; MIR7; PIR-B; PIRB

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UniProt ID: (Human) Q8NHL6

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Entrez Gene ID: (Human) 10859

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Function(s)
protein phosphatase 1 binding HLA-A specific inhibitory MHC class I receptor activity HLA-B specific inhibitory MHC class I receptor activity MHC class I receptor activity SH2 domain binding MHC class I protein binding protein homodimerization activity
Process(es)
negative regulation of T cell mediated cytotoxicity positive regulation of defense response to virus by host adaptive immune response T cell proliferation involved in immune response negative regulation of cytokine secretion involved in immune response immune response-inhibiting cell surface receptor signaling pathway Fc receptor mediated inhibitory signaling pathway signal transduction response to virus positive regulation of gene expression negative regulation of serotonin secretion receptor internalization interferon-gamma production negative regulation of interferon-gamma production negative regulation of mononuclear cell proliferation negative regulation of interferon-beta secretion negative regulation of T cell proliferation negative regulation of tumor necrosis factor biosynthetic process positive regulation of apoptotic process negative regulation of interferon-gamma biosynthetic process negative regulation of cell cycle negative regulation of endocytosis positive regulation of cytolysis positive regulation of transcription from RNA polymerase II promoter negative regulation of natural killer cell mediated cytotoxicity negative regulation of alpha-beta T cell activation regulation of immune response defense response to virus negative regulation of calcium ion transport cellular response to lipopolysaccharide interferon-gamma secretion dendritic cell differentiation negative regulation of dendritic cell apoptotic process negative regulation of interleukin-10 secretion negative regulation of interleukin-12 secretion negative regulation of CD8-positive, alpha-beta T cell activation negative regulation of T cell activation via T cell receptor contact with antigen bound to MHC molecule on antigen presenting cell positive regulation of gamma-delta T cell activation involved in immune response negative regulation of dendritic cell differentiation negative regulation of transforming growth factor-beta secretion negative regulation of osteoclast development
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