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BV421 Mouse Anti-Human CD314 (NKG2D)
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This product is the replacement for 743558.

BV421 Mouse Anti-Human CD314 (NKG2D)

Clone 1D11

(RUO)
BV421 Mouse Anti-Human CD314 (NKG2D)
Two-color flow cytometric analysis of CD314 (NKG2D) expression on human peripheral blood lymphocytes. Human whole blood was stained with PE Anti-Human CD56 (Cat. No. 555516) and with either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; Left Plot) or BD Horizon™ BV421 Mouse Anti-Human CD314 (NKG2D) antibody (Cat. No. 568106; Right Plot) at 0.5 µg/test. The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). A bivariate pseudocolor density plot showing the correlated expression of CD314 (NKG2D) [or Ig Isotype control staining] versus CD56 was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Two-color flow cytometric analysis of CD314 (NKG2D) expression on human peripheral blood lymphocytes. Human whole blood was stained with PE Anti-Human CD56 (Cat. No. 555516) and with either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; Left Plot) or BD Horizon™ BV421 Mouse Anti-Human CD314 (NKG2D) antibody (Cat. No. 568106; Right Plot) at 0.5 µg/test. The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). A bivariate pseudocolor density plot showing the correlated expression of CD314 (NKG2D) [or Ig Isotype control staining] versus CD56 was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Product Details
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BD Horizon™
CD314; KLRK1; KLR; NKG2D; NKG2-D; NK cell receptor D
Human (QC Testing)
Mouse RBF/ DnJ IgG1, κ
NKL cells
Flow cytometry (Routinely Tested)
0.2 mg/ml
VIII 80432
22914
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 to the dye under optimum conditions and unconjugated antibody and free dye were 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® CompBeads are appropriate for your specific cellular application.

For optimal and reproducible results, BD Horizon Brilliant Stain Buffer should be used anytime BD Horizon Brilliant dyes are used in a multicolor flow cytometry panel.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. 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. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. 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.
  3. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  4. 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.
  5. 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.
  6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  7. Pacific Blue™ is a trademark of Life Technologies Corporation.
  8. Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
  9. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  10. An isotype control should be used at the same concentration as the antibody of interest.
568106 Rev. 1
Antibody Details
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1D11

The 1D11 monoclonal antibody specifically binds to NKG2D, a 42 kDa type II transmembrane glycoprotein that is also known as CD314 and KLRK1. NKG2D is a member of the C-type lectin family and is expressed on human NK cells. This activating receptor binds strongly to several ligands including MICA and MICB and ULBP-1, -2, and -3 proteins that are expressed by different target cell types. Different from natural cytotoxicity receptor (NCR), NKG2D expression is not confined to NK cells. It is also expressed on virtually all TCR γ/δ+ and CD8+TCR α/β+ T cells. NKG2D functions as a triggering receptor involved in natural cytotoxicity mediated by normal NK cells against a variety of tumors or normal target cells. Importantly, NKG2D can complement the role of NCR in tumor cell lysis. Remarkably, the combined maskings of NCR and NKG2D can reportedly lead to a complete inhibition of NK-mediated lysis of all tumor or normal cells. The 1D11 antibody can reportedly block or stimulate the function of NKG2D-positive cells.

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.

568106 Rev. 1
Citations & References
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Development References (5)

  1. Bauer S, Groh V, Wu J, et al. Activation of NK cells and T cells by NKG2D, a receptor for stress-inducible MICA. Science. 1999; 285(5428):727-729. (Immunogen: Blocking, Flow cytometry, Functional assay, Immunoprecipitation, Inhibition). View Reference
  2. Groh V, Bruhl A, El-Gabalawy H, Nelson JL, Spies T. Stimulation of T cell autoreactivity by anomalous expression of NKG2D and its MIC ligands in rheumatoid arthritis. Proc Natl Acad Sci U S A. 2003; 100(16):9452-9457. (Clone-specific: Flow cytometry, Immunohistochemistry). View Reference
  3. Groh V, Rhinehart R, Randolph-Habecker J, Topp MS, Riddell SR, Spies T. Costimulation of CD8alphabeta T cells by NKG2D via engagement by MIC induced on virus-infected cells. Nat Immunol. 2001; 2(3):255-260. (Clone-specific: Blocking, (Co)-stimulation, Immunohistochemistry, Inhibition). View Reference
  4. Roberts AI, Lee L, Schwarz E, et al. NKG2D receptors induced by IL-15 costimulate CD28-negative effector CTL in the tissue microenvironment. J Immunol. 2001; 167(10):5527-5530. (Clone-specific: Flow cytometry, Stimulation). View Reference
  5. Steinle A, Li P, Morris DL, et al. Interactions of human NKG2D with its ligands MICA, MICB, and homologs of the mouse RAE-1 protein family. Immunogenetics. 2001; 53(4):279-287. (Clone-specific: Blocking). View Reference
View All (5) View Less
568106 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.