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RB744 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
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 that minimize unconjugated dye and antibody.

Recommended Assay Procedures

BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® 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 BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  3. For U.S. patents that may apply, see bd.com/patents.
  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. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  6. 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.
  7. 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.
  8. When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
  9. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  10. An isotype control should be used at the same concentration as the antibody of interest.
  11. Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
757034 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.

757034 Rev. 1
Format Details
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RB744
The BD Horizon RealBlue™ 744 (RB744) Dye is part of the BD® family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 498-nm and an emission maximum (Em Max) at 746-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RB744 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with minimal excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), we recommend using an optical filter centered near 750-nm (e.g., a 750/60-nm bandpass filter).
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RB744
Blue 488 nm
498 nm
746 nm
757034 Rev.1
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
757034 Rev. 1

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.