BB700 Mouse Anti-Human CD158e1 (NKB1)

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BD OptiBuild™ BB700 Mouse Anti-Human CD158e1 (NKB1)

Name: BB700 Mouse Anti-Human CD158e1 (NKB1)
Brand: BD OptiBuild™
SKU: 745770

Clone DX9

(RUO)

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Scientific Resources

Product Details

Brand: BD OptiBuild™

Alternative Name: KIR3DL1; CD158E; CD158E1; KIR antigen 3DL1; KIR; KI3L1; NKAT-3; NKB1; NKB1B

Reactivity: Human (Tested in Development)

Isotype: Mouse BALB/c IgG1, κ

Immunogen: Human NK Cell Clone

Application: Flow cytometry (Qualified)

Concentration: 0.2 mg/ml

Workshop Number: VII 70319

Entrez Gene ID: 3811

RRID: AB_2743232

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 to the dye under optimum conditions that minimize unconjugated dye and antibody.

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 two washes after staining with antibodies. Cells may be prepared, stained with antibodies and washed twice with wash buffer per established protocols for immunofluorescent 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

The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
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.
BD Horizon Brilliant Blue 700 is covered by one or more of the following US patents: 8,455,613 and 8,575,303.
Cy is a trademark of GE Healthcare.
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

Human BD Fc Block™ RUO

Size 50 µg Cat No. 564219

Stain Buffer (FBS) RUO

Size 500 mL Cat No. 554656

Stain Buffer (BSA) RUO

Size 500 mL Cat No. 554657

Brilliant Stain Buffer RUO

Size 100 Tests Cat No. 563794

Lysing Buffer RUO

Size 100 mL Cat No. 555899

Lysing Solution 10X Concentrate IVD

Size 100 mL Cat No. 349202

View Details

745770 Rev. 1

Antibody Details

DX9

The DX9 monoclonal antibody specifically binds to CD158e1, also known as NKB1. CD158e1 functions as a killer cell inhibitory receptor (KIR) and is encoded by KIR3DL1 (Killer cell immunoglobulin-like receptor, three domains, long cytoplasmic tail). CD158e1 is a 70 kDa glycoprotein that belongs to the Ig superfamily. It is expressed on a subset of natural killer cells and a small subset of T cells. Expression of CD158e1 has been observed to vary among individuals. KIR molecules specifically recognize a certain group of HLA class I antigens. Interaction of CD158e1 with specific HLA-B antigen on a target cell appears to inhibit cell-mediated cytotoxicity by delivering a negative signal that prevents lymphocyte activation. It is suggested that this MHC class I-KIR interaction works as a signaling mechanism that regulates NK and T-cell responses to antigenic challenge.

The antibody was conjugated to BD Horizon BB700, which is part of the BD Horizon Brilliant™ Blue family of dyes. It is a polymer-based tandem dye developed exclusively by BD Biosciences. With an excitation max of 485 nm and an emission max of 693 nm, BD Horizon BB700 can be excited by the 488 nm laser and detected in a standard PerCP-Cy™5.5 set (eg, 695/40-nm filter). This dye provides a much brighter alternative to PerCP-Cy5.5 with less cross laser excitation off the 405 nm and 355 nm lasers.

745770 Rev. 1

Format Details

BB700

The BD Horizon Brilliant™ Blue 700 (BB700) Dye is part of the BD Horizon Brilliant™ Blue family of dyes. This tandem fluorochrome is comprised of a polymer-technology dye donor with an excitation maximum (Ex Max) of 476-nm and an acceptor dye with an emission maximum (Em Max) at 695-nm. Driven by BD innovation, BB700 is designed to be excited by the blue laser (488-nm) and detected using an optical filter centered near 695-nm (e.g., a 695/20-nm bandpass filter). The donor dye can be excited by the Violet (405 nm) laser and the acceptor dye can be excited by the red (627–640 nm) laser resulting in cross-laser excitation and fluorescence spillover. BB700 Reagents are significantly brighter than equivalent PerCP or PerCP-Cy5.5 reagents and are less sensitive to photobleaching. In addition, BB700 shows much less excitation by the violet (407-nm) laser resulting in less spillover. BB700 has minimal yellow green (562-nm) excitation and is ideal for instruments with both blue (488-nm) and yellow green (562-nm) lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.

Format: BB700

Excitation Source: Blue 488 nm

Excitation Max: 476 nm

Emission Max: 695 nm

745770 Rev.1

Citations & References

Development References (10)

D'Andrea A, Chang C, Franz-Bacon K, McClanahan T, Phillips JH, Lanier LL. Molecular cloning of NKB1. A natural killer cell receptor for HLA-B allotypes. J Immunol. 1995; 155(5):2306-2310. (Biology). View Reference
D'Andrea A, Chang C, Phillips JH, Lanier LL. Regulation of T cell lymphokine production by killer cell inhibitory receptor recognition of self HLA class I alleles. J Exp Med. 1996; 184(2):789-794. (Biology). View Reference
Fry AM, Lanier LL, Weiss A. Phosphotyrosines in the killer cell inhibitory receptor motif of NKB1 are required for negative signaling and for association with protein tyrosine phosphatase 1C. J Exp Med. 1996; 184(1):295-300. (Biology). View Reference
Gumperz JE, Paterson JC, Litwin V, et al. Specificity of two anti-class I HLA monoclonal antibodies that block class I recognition by the NKB1 killer cell inhibitory receptor. Tissue Antigens. 1996; 48(4):278-284. (Clone-specific: Blocking, Inhibition). View Reference
Gumperz JE, Valiante NM, Parham P, Lanier LL, Tyan D. Heterogeneous phenotypes of expression of the NKB1 natural killer cell class I receptor among individuals of different human histocompatibility leukocyte antigens types appear genetically regulated, but not linked to major histocompatibililty complex. J Exp Med. 1996; 183(4):1817-1827. (Clone-specific: Blocking, Flow cytometry, Fluorescence activated cell sorting, Functional assay, Immunoprecipitation, Inhibition). View Reference
Lanier LL, Peterson M, Long EO. Antibody reactivity with NK receptors expressed on transfected cells. 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; 2002:413-414.
Litwin V, Gumperz J, Parham P, Phillips JH, Lanier LL. NKB1: a natural killer cell receptor involved in the recognition of polymorphic HLA-B molecules. J Exp Med. 1994; 180(2):537-543. (Immunogen: Bioassay, Blocking, Flow cytometry, Immunoprecipitation, Radioimmunoassay). View Reference
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.
Sivori S, Vitale M, Bottino C, et al. CD94 functions as a natural killer cell inhibitory receptor for different HLA class I alleles: identification of the inhibitory form of CD94 by the use of novel monoclonal antibodies. Eur J Immunol. 1996; 26(10):2487-2492. (Biology). View Reference
Wagtmann N, Rajagopalan S, Winter CC, Peruzzi M, Long EO. Killer cell inhibitory receptors specific for HLA-C and HLA-B identified by direct binding and by functional transfer. Immunity. 1995; 3(6):801-809. (Biology). View Reference

View All (10)

745770 Rev. 1

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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.