Two-color flow cytometric analysis of human ILT4 (CD85d) expression on human peripheral blood monocytes. Whole blood was stained with FITC Mouse Anti-Human CD14 antibody (Cat. No. 347493) and either Alexa Fluor® 647 Mouse IgG2a, κ Isotype Control (Cat. No. 557715; Left Panel) or Alexa Fluor® 647 Mouse Anti-Human ILT4 (CD85d) antibody (Cat. No. 564345; Right Panel). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). Two color flow cytometric contour plots show the correlated expression of ILT4 (CD85d) [or Ig Isotype control staining] versus CD14 for cells with the forward and side light-scatter characteristics of viable monocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Two-color flow cytometric analysis of human ILT4 (CD85d) expression on human peripheral blood monocytes. Whole blood was stained with FITC Mouse Anti-Human CD14 antibody (Cat. No. 347493) and either Alexa Fluor® 647 Mouse IgG2a, κ Isotype Control (Cat. No. 557715; Left Panel) or Alexa Fluor® 647 Mouse Anti-Human ILT4 (CD85d) antibody (Cat. No. 564345; Right Panel). Erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). Two color flow cytometric contour plots show the correlated expression of ILT4 (CD85d) [or Ig Isotype control staining] versus CD14 for cells with the forward and side light-scatter characteristics of viable monocytes. Flow cytometric analysis was performed using a BD LSRFortessa™ Cell Analyzer System.
Product Details
BD Pharmingen™
ILT-4; Ig-like transcript 4; LIR-2; Leukocyte Ig-like receptor 2; MIR10
Human (QC Testing)
Mouse IgG2a, κ
Human CD85d Recombinant Protein
Flow cytometry (Routinely Tested)
5 µl/test
AB_2738758
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO
Preparation And Storage
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.
Product Notices
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
Alexa Fluor® 647 Mouse IgG2a, κ Isotype Control RUO
Size
100 Tests
Cat No.
557715
564345 Rev. 1
Antibody Details
287219
The 287219 monoclonal antibody specifically binds to ILT4 (Immunoglobulin-like transcript 4). ILT-4 is a transmembrane glycoprotein that belongs to the ILT/LIR family within the Ig gene superfamily. It is also known as CD85d, LIR-2 (Leukocyte immunoglobulin-like receptor 2), and MIR-10 (Monocyte/macrophage immunoglobulin-like receptor 10). CD85d is expressed on monocytes, macrophages, and dendritic cells. CD85d binds to HLA class I molecules and to trophoblast-specific HLA-G1. CD85d is reportedly involved in the generation of regulatory T cells and possibly maternal-fetal tolerance.
564345 Rev. 1
Format Details
Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
564345 Rev.1
Citations & References
Development References (5)
-
Allan DS, Colonna M, Lanier LL, et al. Tetrameric complexes of human histocompatibility leukocyte antigen (HLA)-G bind to peripheral blood myelomonocytic cells. J Exp Med. 1999; 189(7):1149-1156. (Biology). View Reference
-
Colonna M, Samaridis J, Cella M, et al. Human myelomonocytic cells express an inhibitory receptor for classical and nonclassical MHC class I molecules. J Immunol. 1998; 160(7):3096-3100. (Biology). View Reference
-
Lichterfeld M, Kavanagh DG, Williams KL, et al. A viral CTL escape mutation leading to immunoglobulin-like transcript 4-mediated functional inhibition of myelomonocytic cells. J Exp Med. 2007; 204(12):2813-2824. (Biology). View Reference
-
Shiroishi M, Kuroki K, Rasubala L, et al. Structural basis for recognition of the nonclassical MHC molecule HLA-G by the leukocyte Ig-like receptor B2 (LILRB2/LIR2/ILT4/CD85d). Proc Natl Acad Sci U S A. 2006; 103(44):16412-16417. (Biology). View Reference
-
Zola H. Leukocyte and stromal cell molecules : the CD markers. Hoboken, N.J.: Wiley-Liss; 2007.
564345 Rev. 1
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.
