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BV421 Mouse Anti-Human CD158a
BV421 Mouse Anti-Human CD158a
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 BD Horizon™ BV421 Mouse IgM, κ Isotype Control (Cat. No. 562704; Left Panel) or BD Horizon BV421 Mouse Anti-Human CD158a antibody (Cat. No. 564318; 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.
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 BD Horizon™ BV421 Mouse IgM, κ Isotype Control (Cat. No. 562704; Left Panel) or BD Horizon BV421 Mouse Anti-Human CD158a antibody (Cat. No. 564318; 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
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BD Horizon™
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_2738741
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 with BD Horizon BV421 under optimum conditions, and unconjugated antibody and free BD Horizon BV421 were removed.

Recommended Assay Procedures

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes (including BD Optibuild Brilliant reagents) are used in the same experiment.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions.  More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

Product Notices

  1. 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).
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. 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.
  4. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  5. Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
  8. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
  9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
564318 Rev. 2
Antibody Details
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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.

The antibody was conjugated to BD Horizon BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.

564318 Rev. 2
Format Details
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BV421
The BD Horizon Brilliant Violet™ 421 (BV421) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology based dye has an excitation maximum (Ex Max) of 407-nm and an emission maximum (Em Max) at 423-nm. Driven by BD innovation, BV421 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 420-nm (e.g., a 431/28-nm or 450/50-nm bandpass filter). BV421 is an ideal alternative for V450 as it is approximately ten times brighter with less spillover into the BV510/V500 detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV421
Violet 405 nm
407 nm
423 nm
564318 Rev.2
Citations & References
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Development References (2)

  1. 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.
  2. 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
564318 Rev. 2

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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.