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BB700 Mouse Anti-Mouse H-2Ld/H-2Db
Product Details
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BD OptiBuild™
H-2Db/H-2Ld; H2-Db/H2-Ld; Histocompatibility-2 Db/Ld
Mouse (Tested in Development)
Mouse C3H, also known as C3H/He, C3H/Bi IgG2a, κ
C3H.SW mouse splenocytes
Flow cytometry (Qualified)
0.2 mg/ml
14964
AB_2743287
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 BB700 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 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 or 566349).

When setting up compensation, it is recommended to compare spillover values obtained from cells and BD™ CompBeads to ensure that beads will provide sufficiently accurate spillover values.

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

  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 Blue 700 is covered by one or more of the following US patents: 8,455,613 and 8,575,303.
  10. Cy is a trademark of GE Healthcare.
745842 Rev. 1
Antibody Details
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28-14-8

The 28-14-8 antibody reacts with the α3 domain of the H-2D[b] MHC class I alloantigen.  The antibody binds to H-2D[b] in the presence or absence of the β2 microglobulin chain.  It cross-reacts with the α3 domain of H-2L[d], but not with K[d] or D[d], and with H-2D[q] and/or L[q].  Reactivity with haplotypes k, f, p, r, and s has not been observed.  mAb 28-14-8 has been shown to block H-2L[d]- specific and H- 2L[d]-restricted antigen-specific lysis of target cells by cytotoxic T lymphocytes (CTL),  but it does not block recognition of H-2L[d] positive target cells by Ly-6G2-positive NK cells.

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.

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Format Details
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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.
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BB700
Blue 488 nm
476 nm
695 nm
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Citations & References
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Development References (9)

  1. Allen H, Fraser J, Flyer D, Calvin S, Flavell R. Beta 2-microglobulin is not required for cell surface expression of the murine class I histocompatibility antigen H-2Db or of a truncated H-2Db. Proc Natl Acad Sci U S A. 1986; 83(19):7447-7451. (Clone-specific: Immunoprecipitation). View Reference
  2. Allen H, Wraith D, Pala P, Askonas B, Flavell RA. Domain interactions of H-2 class I antigens alter cytotoxic T-cell recognition sites. Nature. 1984; 309(5965):279-281. (Clone-specific: Radioimmunoassay). View Reference
  3. Evans GA, Margulies DH, Shykind B, Seidman JG, Ozato K. Exon shuffling: mapping polymorphic determinants on hybrid mouse transplantation antigens. Nature. 1982; 300(5894):755-757. (Clone-specific: Radioimmunoassay). View Reference
  4. Kündig TM, Bachmann MF, DiPaolo C. Fibroblasts as efficient antigen-presenting cells in lymphoid organs. Science. 1995; 268(5215):1343-1347. (Clone-specific: Blocking). View Reference
  5. Mason LH, Ortaldo JR, Young HA, Kumar V, Bennett M, Anderson SK. Cloning and functional characteristics of murine large granular lymphocyte-1: a member of the Ly-49 gene family (Ly-49G2). J Exp Med. 1995; 182(2):293-303. (Clone-specific: Blocking). View Reference
  6. Orn A, Goodenow RS, Hood L. Product of a transferred H-2Ld gene acts as restriction element for LCMV-specific killer T cells. Nature. 1982; 297(5865):415. (Clone-specific: Blocking). View Reference
  7. Ozato K, Hansen TH, Sachs DH. Monoclonal antibodies to mouse MHC antigens. II. Antibodies to the H-2Ld antigen, the products of a third polymorphic locus of the mouse major histocompatibility complex. J Immunol. 1980; 125(6):2473-2477. (Immunogen: Cytotoxicity). View Reference
  8. Ozato K, Sachs DH. Monoclonal antibodies to mouse MHC antigens. III. Hybridoma antibodies reacting to antigens of the H-2b haplotype reveal genetic control of isotype expression. J Immunol. 1981; 126(1):317-321. (Immunogen: Cytotoxicity). View Reference
  9. Woodward JG, Orn A, Harmon RC, Goodenow RS, Hood L, Frelinger JA. Specific recognition of the product of a transferred major histocompatibility complex gene by cytotoxic T lymphocytes. Proc Natl Acad Sci U S A. 1982; 79(11):3613-3617. (Clone-specific: Blocking). View Reference
View All (9) View Less
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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.