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BUV661 Rat Anti-Mouse H-2 Class I
Product Details
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BD OptiBuild™
H-2 Class I; H2 Class I; Mouse MHC Class I; Mouse MHC-I
Mouse (Tested in Development)
Rat DA, also known as DA/HA IgG2a, κ
Mouse B10 Spleen Cells
Flow cytometry (Qualified)
0.2 mg/ml
AB_2873956
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 BUV661 under optimal conditions that minimize unconjugated dye and antibody.

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).

Product Notices

  1. This antibody was developed for use in flow cytometry.
  2. 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.
  3. Researchers should determine the optimal concentration of this reagent for their individual applications.
  4. An isotype control should be used at the same concentration as the antibody of interest.
  5. 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.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  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. BD Horizon Brilliant Ultraviolet 661 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,575,303; 8,354,239.
749702 Rev. 4
Antibody Details
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M1/42

The M1/42 monoclonal antibody (mAb) specifically recognizes intact Histocompatibility-2 Region (H-2) class I antigens (Ags), also known as mouse Major Histocompatibility Complex class I (MHC-I) Ags. This mAb reportedly binds to one or more of the H-2 Ags (H-2K, H-2D, and likely H-2L) that are primarily expressed on most nucleated somatic cells of hematopoietic and nonhematopoietic origin. H-2 class I molecules are comprised of a ~45-50 kDa type I transmembrane heavy chain glycoprotein that is noncovalently linked to ~13 kDa β2-microglobulin (β2-m). The extracellular region of the polymorphic heavy chains is comprised of three globular domains (α1, α2, and α3), followed by a transmembrane region and a cytoplasmic tail. Sequence variations are manifest in regions of the α1 and α2 domains that line the peptide-binding cleft involved in Ag presentation. The M1/42 mAb reportedly binds to H-2 class I Ags from multiple mouse haplotypes including a, b, d, j, k, s, and u. It does not bind to separated H-2 class I heavy chains or β2-m. Cell surface CD8 molecules bind to invariant sites of the H-2 heavy chains and can provide coreceptor signaling for Ag-mediated activation through the T cell receptor. H-2 class I molecules that present self-Ags can lead to self-tolerance of maturing CD8+ T cells due to thymic selection. Alternatively, these molecules can present foreign peptide Ags to mature peripheral CD8+ T cells resulting in cell-mediated immune responses against foreign Ags. H-2 class I Ags also serve as ligands for activating and inhibitory receptors expressed by NK cells and some T cells. The M1/42 antibody is useful for analyzing H-2 class I Ag expression on cells or cell lines. Cells from different experimental systems, eg, stressed cells that have undergone infection or transformation, may express little or no H-2 class I Ag. In contrast, cells undergoing activation or responding to certain factors (eg, interferons) may express upregulated levels of these Ags.

The antibody was conjugated to BD Horizon™ BUV661 which is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This dye is a tandem fluorochrome of BD Horizon BUV395 with an Ex Max of 348-nm and an acceptor dye with an Em Max at 661-nm. BD Horizon Brilliant BUV661 can be excited by the ultraviolet laser (355 nm) and detected with a 670/25 filter and a 630 nm LP.  Due to cross laser excitation of this dye, there may be significant spillover into channels detecting APC-like emissions (eg, 670/25-nm filter).

    

Due to spectral differences between labeled cells and beads, using BD™ CompBeads can result in incorrect spillover values when used with BD Horizon BUV661 reagents. Therefore, the use of BD CompBeads or BD CompBeads Plus to determine spillover values for these reagents is not recommended. Different BUV661 reagents (eg, CD4 vs. CD45) can have slightly different fluorescence spillover therefore, it may also be necessary to use clone-specific compensation controls when using these reagents.

749702 Rev. 4
Format Details
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BUV661
The BD Horizon Brilliant™ Ultraviolet 661 (BUV661) Dye is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This tandem fluorochrome is comprised of a BUV395 donor with an excitation maximum (Ex Max) of 350-nm and an acceptor dye with an emission maximum (Em Max) at 660-nm. BUV661, driven by BD innovation, is designed to be excited by the ultraviolet laser (355-nm) and detected using an optical filter centered near 660-nm (e.g., 670/25 bandpass filter). The acceptor dye can be excited by the Red (628–640-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BUV661
Ultraviolet 355 nm
350 nm
660 nm
749702 Rev.4
Citations & References
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Development References (8)

  1. Cornish AL, Freeman S, Forbes G, et al. Characterization of siglec-5, a novel glycoprotein expressed on myeloid cells related to CD33. Blood. 1998; 92(6):2123-2132. (Immunogen: ELISA, Flow cytometry). View Reference
  2. Crocker PR, Varki A. Siglecs, sialic acids and innate immunity. Trends Immunol. 2001; 22(6):337-342. (Biology). View Reference
  3. Jones C, Virji M, Crocker PR. Recognition of sialylated meningococcal lipopolysaccharide by siglecs expressed on myeloid cells leads to enhanced bacterial uptake.. Mol Microbiol. 2003; 49(5):1213-25. (Clone-specific: Blocking, Functional assay). View Reference
  4. Nguyen DH, Hurtado-Ziola N, Gagneux P, Varki A. Loss of Siglec expression on T lymphocytes during human evolution. Proc Natl Acad Sci U S A. 2006; 103(20):7765-7770. (Clone-specific: Flow cytometry). View Reference
  5. Pillai S, Netravali IA, Cariappa A, Mattoo H. Siglecs and immune regulation.. Annu Rev Immunol. 2012; 30:357-92. (Biology). View Reference
  6. Simmons DL, Buckley CD, Schneemann M. Adhesion Structures: Section 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; 2002:3-14.
  7. Zola H, Swart B, Boumsell L, Mason DY. Human Leucocyte Differentiation Antigen nomenclature: update on CD nomenclature. Report of IUIS/WHO Subcommittee.. J Immunol Methods. 2003; 275(1-2):1-8. (Clone-specific). View Reference
  8. Zola H, Swart B, Nicholson I, Voss E. CD170. In: Zola H. Leukocyte and Stromal Cell Molecules : the CD Markers. Hoboken, N.J.: Wiley-Liss; 2007:309-310.
View All (8) View Less
749702 Rev. 4

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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.