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BV421 Mouse Anti-Mouse H-2Kd/H-2Dd
BV421 Mouse Anti-Mouse H-2Kd/H-2Dd

Flow cytometric analysis of H-2Kd/H-2Dd expression on BALB/c mouse splenocytes. Splenic leucocytes from a C3H (H-2k haplotype; Left Plot) or a BALB/c (H-2d haplotype; Right Plot) mouse were stained with BD Horizon™ BV421 Mouse IgG2a, κ Isotype Control (Cat. No. 562439; dashed line histograms) or with BD Horizon™ BV421 Mouse Anti-Mouse H-2Kd/H-2Dd antibody (Cat. No. 567294; solid line histograms) at 0.25 µg/test. The fluorescence histograms showing H-2Kd/H-2Dd expression (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of viable splenic leucocytes. 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.

Flow cytometric analysis of H-2Kd/H-2Dd expression on BALB/c mouse splenocytes. Splenic leucocytes from a C3H (H-2k haplotype; Left Plot) or a BALB/c (H-2d haplotype; Right Plot) mouse were stained with BD Horizon™ BV421 Mouse IgG2a, κ Isotype Control (Cat. No. 562439; dashed line histograms) or with BD Horizon™ BV421 Mouse Anti-Mouse H-2Kd/H-2Dd antibody (Cat. No. 567294; solid line histograms) at 0.25 µg/test. The fluorescence histograms showing H-2Kd/H-2Dd expression (or Ig Isotype control staining) were derived from gated events with the forward and side light-scatter characteristics of viable splenic leucocytes. 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™
H2-D1, H-2D; H2-K1, H-2K, H2-K
Mouse (QC Testing)
Mouse C3H, also known as C3H/He, C3H/Bi IgG2a, κ
C57BL/6 × DBA/2 (BDF1) splenocytes
Flow cytometry (Routinely Tested)
0.2 mg/ml
Aqueous buffered solution containing BSA and ≤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 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).

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  4. 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.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. 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.
  7. 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.
  8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  9. Pacific Blue™ is a trademark of Life Technologies Corporation.
  10. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
567294 Rev. 2
Antibody Details
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34-1-2S

The 34-1-2S monoclonal antibody specifically recognizes the mouse histocompatibility 2 (H-2) alloantigens, H-2Kd and H-2Dd, that are variably expressed on most nucleated cells. These MHC class I antigens are heterodimers comprised of a polymorphic alpha heavy chains (~44 kDa type I transmembrane glycoprotein encoded by the H-2 gene complex) that are noncovalently associated with invariant β2-microglobulin (~11 kDa beta light chain encoded by B2m). H-2Kd and H-2Dd are involved in the positive and negative selection of CD8+ T cells in the thymus as well as MHC-restricted antigen presentation to CD8+ αβ T cells in the periphery. H-2Kd and H-2Dd molecules can also serve a ligands for activating or inhibitory receptors, such as, those encoded by the Ly49 gene family which are variably expressed on subsets of natural killer (NK) cells and T cells. The 34-1-2S antibody cross-reacts with H-2K MHC class I alloantigens of the b, s, r, q, or p haplotypes.

The antibody was conjugated to BD Horizon BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max near 407 nm and Em Max near 421 nm, BD Horizon BV421 can be excited by the violet laser (405 nm) and detected with a 450/50 nm filter. BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue™ conjugates. Due to nearly identical excitation and emission properties but different spillover characteristics, BD Horizon BV421, Pacific Blue™, and BD Horizon V450 cannot be used simultaneously.

567294 Rev. 2
Format Details
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BV421
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.
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BV421
Violet 405 nm
407 nm
423 nm
567294 Rev.2
Citations & References
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Development References (6)

  1. Boudreau JE, Hsu KC. Natural Killer Cell Education and the Response to Infection and Cancer Therapy: Stay Tuned.. Trends Immunol. 2018; 39(3):222-239. (Methodology). View Reference
  2. Brennan J, Mager D, Jefferies W, Takei F. Expression of different members of the Ly-49 gene family defines distinct natural killer cell subsets and cell adhesion properties. J Exp Med. 1994; 180(6):2287-2295. (Clone-specific: Blocking). View Reference
  3. Lenz A, Heufler C, Rammensee HG, et al. Murine epidermal Langerhans cells express significant amounts of class I major histocompatibility complex antigens.. Proc Natl Acad Sci U S A. 1989; 86(19):7527-31. (Clone-specific: Flow cytometry, Immunofluorescence). View Reference
  4. Ozato K, Mayer NM, Sachs DH. Monoclonal antibodies to mouse major histocompatibility complex antigens IV. A series of hybridoma clones producing anti-H-2d antibodies and an examination of expression of H-2d antigens on the surface of these cells. Transplantation. 1982; 34(3):113-120. (Immunogen: Cytotoxicity, Radioimmunoassay). View Reference
  5. Pereira RA, Simmons A. Cell surface expression of H2 antigens on primary sensory neurons in response to acute but not latent herpes simplex virus infection in vivo.. J Virol. 1999; 73(8):6484-9. (Clone-specific: Flow cytometry). View Reference
  6. Schenkel AR, Kingry LC, Slayden RA. The ly49 gene family. A brief guide to the nomenclature, genetics, and role in intracellular infection.. Front Immunol. 2013; 4:90. (Biology). View Reference
View All (6) View Less
567294 Rev. 2

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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.

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.