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RB780 Hamster Anti-Mouse CD27
RB780 Hamster Anti-Mouse CD27
Multiparameter flow cytometric analysis using BD OptiBuild™ RB780 Hamster Anti-Mouse CD27 antibody (Cat. No. 755925; Right Plot) on viable C57BL/6 mouse splenocytes, with Autofluorescence Control (Left Plot). Flow cytometry was performed using a BD LSRFortessa™ X-20 Flow Cytometer System.
Multiparameter flow cytometric analysis using BD OptiBuild™ RB780 Hamster Anti-Mouse CD27 antibody (Cat. No. 755925; Right Plot) on viable C57BL/6 mouse splenocytes, with Autofluorescence Control (Left Plot). Flow cytometry was performed using a BD LSRFortessa™ X-20 Flow Cytometer System.
Product Details
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BD OptiBuild™
Tnfrsf7; Tumor necrosis factor receptor superfamily member 7; Tp55; S152
Mouse (Tested in Development)
Armenian Hamster IgG1, κ
Armenian hamster fibroblast line ARHO12 transfected with mouse Cd27 cDNA
Flow cytometry (Qualified)
0.2 mg/ml
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. 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.
  2. Researchers should determine the optimal concentration of this reagent for their individual applications.
  3. 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.
  4. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  5. Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
  6. An isotype control should be used at the same concentration as the antibody of interest.
  7. 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.
  8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  9. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  10. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  11. Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
755925 Rev. 2
Antibody Details
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LG.3A10

The LG.3A10 monoclonal antibody specifically binds to CD27, a lymphocyte-restricted member of the Tumor Necrosis Factor Receptor family which binds to CD70. The CD27 molecule is a 45-kDa transmembrane glycoprotein which is constitutively expressed by lymphocytes of the T lineage: virtually all thymocytes and over 90% of peripheral T cells bearing both αβ and γδ T-cell receptors. CD27 cooperates with the pre-TCR in mediating thymocyte differentiation and expansion. In addition, one to ten percent of mature peripheral B cells express CD27, and CD27's role in the differentiation of human plasma cells has been studied. Mouse NK cells, freshly isolated and IL-2-activated, also express CD27. In the bone marrow, CD27 is found on a progenitor population which provides short-term hematopoietic reconstitution. Cells of the myeloid lineage do not express CD27. Cross-linked LG.3A10 mAb has been reported to amplify the proliferative response of purified T lymphocytes to suboptimal mitogenic stimulation1 and to enhance NK-cell proliferation and IFN-γ production. In contrast, non-cross-linked LG.3A10 mAb inhibits CD3-induced pre-T cell development by interfering with the receptor-ligand interaction. This hamster mAb to a mouse leukocyte antigen has been observed to cross-react with a similar population of rat leukocytes.

755925 Rev. 2
Format Details
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RB780
The BD Horizon RealBlue™ 780 (RB780) 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 781-nm. Driven by BD innovation, RB780 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), RB780 can be used as an alternative to PE-Cy7 and we recommend using an optical filter centered near 780-nm (eg, a 780/60-nm bandpass filter). For spectral instruments equipped with a Blue laser (488-nm), it can be used in conjunction with PE-Cy7. RB780 is on average brighter than PE-Cy7 and has minimal spillover into Yellow-Green detectors.
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RB780
Blue 488 nm
498 nm
781 nm
755925 Rev.2
Citations & References
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View product citations for antibody "755925" on CiteAb

Development References (6)

  1. Agematsu K, Hokibara S, Nagumo H, Shinozaki K, Yamada S, Komiyama A. Plasma cell generation from B-lymphocytes via CD27/CD70 interaction. Leuk Lymphoma. 1999; 35(3-4):219-225. (Biology). View Reference
  2. Gravestein LA, Blom B, Nolten LA, et al. Cloning and expression of murine CD27: comparison with 4-1BB, another lymphocyte-specific member of the nerve growth factor receptor family. Eur J Immunol. 1993; 23(4):943-950. (Biology). View Reference
  3. Gravestein LA, Nieland JD, Kruisbeek AM, Borst J. Novel mAbs reveal potent co-stimulatory activity of murine CD27. Int Immunol. 1995; 7(4):551-557. (Immunogen: (Co)-stimulation, Flow cytometry, Functional assay, Immunofluorescence, Immunoprecipitation). View Reference
  4. Gravestein LA, van Ewijk W, Ossendorp F, Borst J. CD27 cooperates with the pre-T cell receptor in the regulation of murine T cell development. J Exp Med. 1996; 184(2):675-685. (Clone-specific: Flow cytometry, Immunofluorescence, Inhibition). View Reference
  5. Takeda K, Oshima H, Hayakawa Y, et al. CD27-mediated activation of murine NK cells. J Immunol. 2000; 164(4):1741-1745. (Clone-specific: (Co)-stimulation, Enhancement, Flow cytometry). View Reference
  6. Wiesmann A, Phillips RL, Mojica M, et al. Expression of CD27 on murine hematopoietic stem and progenitor cells. Immunity. 2000; 12(2):193-199. (Clone-specific: Flow cytometry). View Reference
View All (6) View Less
755925 Rev. 2

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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.