Two-color flow cytometric analysis of CD158a expression on human peripheral blood lymphocytes. Whole blood was stained with PE Mouse Anti-Human CD16 antibody (Cat. No. 555407/560995) and either APC Mouse IgM, κ Isotype Control (Cat. No. 550883; Left Panel) or APC Mouse Anti-Human CD158a antibody (Cat. No. 564319; Right Panel). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). Two-color flow cytometric contour plots showing the correlated expression of CD158a (or Ig Isotype control staining) versus CD16 were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
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Two-color flow cytometric analysis of CD158a expression on human peripheral blood lymphocytes. Whole blood was stained with PE Mouse Anti-Human CD16 antibody (Cat. No. 555407/560995) and either APC Mouse IgM, κ Isotype Control (Cat. No. 550883; Left Panel) or APC Mouse Anti-Human CD158a antibody (Cat. No. 564319; Right Panel). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). Two-color flow cytometric contour plots showing the correlated expression of CD158a (or Ig Isotype control staining) versus CD16 were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Product Details
BD Pharmingen™
KIR2DL1; KI2L1; NKAT; NKAT-1; NKAT1; KIR-K64; KIR221; p58.1
Human (QC Testing)
Mouse BALB/c IgM
Human NK Cells
Flow cytometry (Routinely Tested)
5 µl
VI BP373, NKNK14
AB_2738742
Aqueous buffered solution containing BSA, protein stabilizer, 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 APC under optimum conditions, and unconjugated antibody and free APC were 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).
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- This APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNe, or red diode laser.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Lysing Solution 10X Concentrate IVD
Size
100 mL
Cat No.
349202
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
APC Mouse IgM, κ Isotype Control RUO
Size
0.1 mg
Cat No.
550883
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PE Mouse Anti-Human CD16 RUO
Size
25 Tests
Cat No.
560995
564319 Rev. 2
Antibody Details
HP-3E4
The HP-3E4 monoclonal antibody recognizes CD158a. CD158a is a 58 kDa membrane glycoprotein, member of the killer-inhibitory receptor (KIR). The KIR family consists of transmembrane glycoproteins of the Ig superfamily expressed on <1-8% of NK cells and a subset of T cells which are involved in recognition of MHC class I molecules on target cells and inhibit cytotoxicity. CD158a is an antigen which recognizes HLA-C molecules (-Cw*0401 and -Cw*1503). This antibody is suitable for staining acetone-fixed, frozen tissue sections. It was reported that this clone also crossreacted with NKAT8 (CD158i) transfected cells.
564319 Rev. 2
Format Details
APC
Allophycocyanin (APC), is part of the BD family of phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 651 nm and an emission maximum (Em Max) at 660 nm. APC is designed to be excited by the Red (627-640 nm) laser and detected using an optical filter centered near 660 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.
APC
Red 627-640 nm
651 nm
660 nm
564319 Rev.2
Citations & References
Development References (3)
-
De Miguel M, López-Botet M. Characterization of monoclonal antibodies specific for receptors of the KIR family. Inmunologia. 2002; 21(4):187-193. (Clone-specific: Flow cytometry). View Reference
-
Kishimoto T. Tadamitsu Kishimoto .. et al., ed. Leucocyte typing VI : white cell differentiation antigens : proceedings of the sixth international workshop and conference held in Kobe, Japan, 10-14 November 1996. New York: Garland Pub.; 1997.
-
Melero I, Salmeron A, Balboa MA, Aramburu J, Lopez-Botet M. Tyrosine kinase-dependent activation of human NK cell functions upon stimulation through a 58-kDa surface antigen selectively expressed on discrete subsets of NK cells and T lymphocytes. J Immunol. 1994; 152(4):1662-1673. (Immunogen: Cytotoxicity, Flow cytometry, Functional assay, Immunofluorescence, Immunohistochemistry, Immunoprecipitation, Stimulation). View Reference
564319 Rev. 2
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.
