BB515 Mouse Anti-Human CD158b

BD Horizon™ BB515 Mouse Anti-Human CD158b

Name: BB515 Mouse Anti-Human CD158b
Brand: BD Horizon™
SKU: 566053

Clone CH-L

(RUO)

Two-color flow cytometric analysis of CD158b expression on human peripheral blood lymphocytes - Staining comparisons between BD Horizon™ BB515- and FITC-conjugated antibodies. Human whole blood was stained with APC Mouse Anti-Human CD56 antibody (Cat. No. 555518) and either BD Horizon BB515 Mouse IgG2b, κ Isotype Control (Cat. No. 564510; Left Panel), BD Horizon BB515 Mouse Anti-Human CD158b antibody (Cat. No. 564678/566053; Middle Panel), or FITC Mouse Anti-Human CD158b antibody (Cat. No. 559784; Right Panel). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No.349202). Two-color flow cytometric contour plots showing the correlated expression of CD158b (or Ig Isotype control staining) versus CD56 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.

Spectrum Viewer

Protocol Library

Scientific Resources

Product Details

Brand: BD Horizon™

Alternative Name: CD158b1/KIR2DL2/NKAT-6; CD158b2/KIR2DL3/NKAT-2; CD158j/KIR2DS2/NKAT-5

Reactivity: Human (QC Testing)

Isotype: Mouse BALB/c IgG2b, κ

Immunogen: Human NK Cells

Application: Flow cytometry (Routinely Tested)

Vol. Per Test: 5 µl

Workshop Number: VI NK8

RRID: AB_2738888

Storage Buffer: Aqueous buffered solution containing ≤0.09% sodium azide.

Regulatory Status: 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™ BB515 under optimum conditions and unconjugated antibody was removed.

Recommended Assay Procedures

BD™ CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime two or more BD Horizon Brilliant dyes 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).

For optimal results, it is recommended to perform 2 washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescence staining, prior to acquisition on a flow cytometer. Performing fewer than the recommended wash steps may lead to increased spread of the negative population.

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.
For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
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.
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

Brilliant Stain Buffer RUO

Size 100 Tests Cat No. 563794

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

BB515 Mouse IgG2b, κ Isotype Control RUO

Size 50 µg Cat No. 564510

View Details

566053 Rev. 1

Antibody Details

CH-L

The CH-L monoclonal antibody specifically binds to CD158b proteins. These proteins are 50-58 kDa type I glycoproteins that belong to the Killer cell immunoglobulin-like receptor (KIR) family: (KIR2DL2/L3/S2). They are also known as CD158b1 (KIR2DL2; NKAT-6; p58.2), CD158b2 (KIR2DL3; NKAT-2; p58.2), or CD158j (KIR2DS2; NKAT-5; p50.2). The CD158b molecules are composed of two extracellular Ig-like domains, and a transmembrane region. CD158b1 and CD158b2 also possess long (84 or 76 amino acids, respectively) cytoplasmic tails with two immunoreceptor tyrosine-based inhibition motifs (ITIM) whereas CD158j has a short (39 amino acid) cytoplasmic tail that lacks the ITIM motif. CD158b molecules are expressed on NK cells and subsets of TCR αβ+ cells or TCR γδ+ cells. Ligand- or CH-L antibody-bound CD158b1 or CD158b2 can reportedly inhibit cytolytic NK and T cell responses to various stimuli including certain target cells expressing MHC class I ligands encoded by HLA-C alleles (Cw 1, 3, 7 and 8). CD158j reportedly can enhance some cellular cytolytic responses.

The antibody was conjugated to BD Horizon BB515 which is part of the BD Horizon Brilliant™ Blue family of dyes. With an Ex Max near 490 nm and an Em Max near 515 nm, BD Horizon BB515 can be excited by the blue laser (488 nm) laser and detected with a 530/30 nm filter. This dye has been exclusively developed by BD Biosciences and is up to seven times brighter than FITC with less spillover into the PE channel. Due to similar excitation and emission properties, BB515, FITC, and Alexa Fluor® 488 cannot be used simultaneously. It is not recommended to use BB515 in cocktails that include Streptavidin conjugates as it may cause high background.

566053 Rev. 1

Format Details

BB515

The BD Horizon Brilliant™ Blue 515 (BB515) dye is part of the BD Horizon Brilliant™ Blue family of dyes. This dye is a polymer fluorochrome with an excitation maximum (Ex Max) at 490-nm and an emission maximum (Em Max) of 515-nm. Driven by BD innovation, BB515 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 520-nm (e.g., 530/30-nm). BB515 reagents are significantly brighter than equivalent FITC or Alexa Fluor™ 488 reagents with less spillover into the PE detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.

Format: BB515

Excitation Source: Blue 488 nm

Excitation Max: 490 nm

Emission Max: 515 nm

566053 Rev.1

Citations & References

Development References (8)

Cambiaggi A, Orengo AM, Meazza R, et al. The natural killer-related receptor for HLA-C expressed on T cells from CD3+ lymphoproliferative disease of granular lymphocytes displays either inhibitory or stimulatory function. Blood. 1996; 87(6):2369-2375. (Biology). View Reference
Colonna M, Samaridis J. Cloning of immunoglobulin-superfamily members associated with HLA-C and HLA-B recognition by human natural killer cells. Science. 1995; 268(5209):405-408. (Biology). View Reference
Ferrini S, Cambiaggi A, Meazza R, et al. T cell clones expressing the natural killer cell-related p58 receptor molecule display heterogeneity in phenotypic properties and p58 function. Eur J Immunol. 1994; 24(10):2294-2298. (Clone-specific: Blocking, Cell separation, Flow cytometry, Functional assay, Inhibition, Radioimmunoassay, Western blot). View Reference
Kim J, Chwae YJ, Kim MY, Choi IH, Park JH, Kim SJ. Molecular basis of HLA-C recognition by p58 natural killer cell inhibitory receptors. J Immunol. 1997; 159(8):3875-3882. (Biology). View Reference
Moretta A, Bottino C, Biassoni R. CD158a (p58.1/p50.1) and CD158b (p58.2/p50.2) natural killer receptors for HLA-C alleles: Workshop Report. In: 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.; 1997:290-292.
Pascal V, Vivier E, Andre P. CD158 (killer immunoglobulin-like receptors family) report. In: Mason D. David Mason .. et al., ed. Leucocyte typing VII : white cell differentiation antigens : proceedings of the Seventh International Workshop and Conference held in Harrogate, United Kingdom. Oxford: Oxford University Press; 2002:412-413.
Warren HS, Kinnear BF. CD158a and b Workshop: Killer-inhibitory receptors and natural killer cell proliferation. In: 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.; 1997:292-294.
van Bergen J, Thompson A, van der Slik A, Ottenhoff TH, Gussekloo J, Koning F. Phenotypic and functional characterization of CD4 T cells expressing killer Ig-like receptors. J Immunol. 2004; 173(11):6719-6726. (Clone-specific: Flow cytometry). View Reference

View All (8)

566053 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.