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BD Horizon™ BB515 Mouse Anti-Human NKp44 (CD336)
Clone p44-8 (RUO)
Multicolor flow cytometric analysis of NKp44 (CD336) expression by IL-2-activated large granular lymphocytes. Human peripheral blood mononuclear cells were cultured (7 days, 37°C) with recombinant human IL-2 protein (Cat. No. 554603). The IL-2-activated cells were then stained with PE Mouse Anti-Human CD3 (Cat. No. 555333/561808/561809) and APC Mouse Anti-Human CD56 (Cat. No. 555518) antibodies, and either BD Horizon™ BB515 Mouse IgG1, κ Isotype Control (Cat. No. 564416; dashed line histogram) or BD Horizon BB515 Mouse Anti-Human NKp44 (CD336) antibody (Cat. No. 565099/566025; solid line histogram). The fluorescence histogram showing NKp44 (CD336) expression (or Ig Isotype control staining) was derived from CD3- CD56+ gated events with the forward and side light-scatter characteristics of large granular lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Multicolor flow cytometric analysis of NKp44 (CD336) expression by IL-2-activated large granular lymphocytes. Human peripheral blood mononuclear cells were cultured (7 days, 37°C) with recombinant human IL-2 protein (Cat. No. 554603). The IL-2-activated cells were then stained with PE Mouse Anti-Human CD3 (Cat. No. 555333/561808/561809) and APC Mouse Anti-Human CD56 (Cat. No. 555518) antibodies, and either BD Horizon™ BB515 Mouse IgG1, κ Isotype Control (Cat. No. 564416; dashed line histogram) or BD Horizon BB515 Mouse Anti-Human NKp44 (CD336) antibody (Cat. No. 565099/566025; solid line histogram). The fluorescence histogram showing NKp44 (CD336) expression (or Ig Isotype control staining) was derived from CD3- CD56+ gated events with the forward and side light-scatter characteristics of large granular lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
Product Details
BD Horizon™
CD336; NCR2; NCTR2; NKp44; NK-p44; LY95; dJ149M18.1
Human (QC Testing)
Mouse BALB/c IgG1, κ
Human NKp44
Flow cytometry (Routinely Tested)
5 µl
AB_2739067
Aqueous buffered solution containing ≤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™ 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
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
BB515 Mouse IgG1, κ Isotype Control RUO
Size
100 µg
Cat No.
564416
PE Mouse Anti-Human CD3 RUO
Size
100 Tests
Cat No.
555333
PE Mouse Anti-Human CD3 RUO
Size
25 Tests
Cat No.
561808
565099 Rev. 2
Antibody Details
p44-8
The p44-8 monoclonal antibody specifically binds to the natural killer (NK) cell receptor, NKp44, which is also known as CD336, Natural cytotoxicity triggering receptor 2 (NCR2), or Lymphocyte antigen 95 homolog (Ly95). NKp44 is a ~44 kDa type I transmembrane protein that belongs to the natural cytotoxicity receptor (NCR) family within the immunoglobulin superfamily. NKp44 is expressed by activated NK cells. NKp44 serves as an activating receptor that can enhance NK cell mediated lysis of target cells including tumor cells and virus-infected cells. Killer activating receptor associated protein (KARAP), which is also known as DAP12, is an intracellular adaptor protein that can associate with the intracellular region of NKp44. DAP12 can then function to help transmit activating signals through its immunoreceptor tyrosine-based activation motif (ITAM).
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.
565099 Rev. 2
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. It is not recommended to use BB515 in cocktails that include streptavidin conjugates as it may cause high background.
BB515
490 nm
515 nm
565099 Rev.2
Citations & References
View product citations for antibody "565099" on CiteAb
Development References (4)
-
Byrd A, Hoffmann SC, Jarahian M, Momburg F, Watzl C. Expression analysis of the ligands for the Natural Killer cell receptors NKp30 and NKp44. PLoS ONE. 2007; 2(12):e1339. (Immunogen: ELISA, Flow cytometry). View Reference
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Lanier LL. Natural killer cell receptor signaling. Curr Opin Immunol. 2003; 15:308-314. (Biology). View Reference
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Moretta L, Moretta A. Unravelling natural killer cell function: triggering and inhibitory human NK receptors. EMBO J. 2004; 23(2):255-259. (Biology). View Reference
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Vitale M, Bottino C, Sivori S, et al. NKp44, a novel triggering surface molecule specifically expressed by activated natural killer cells, is involved in non-major histocompatibility complex-restricted tumor cell lysis.. J Exp Med. 187(12):2065-2072. (Biology). View Reference
565099 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.
Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product 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.