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BUV661 Mouse Anti-Human CD148
Product Details
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BD OptiBuild™
DEP-1; DEP1; HPTP eta; HPTP-η; PTPRJ; R-PTP-eta; R-PTP-J; SCC1; p260
Human (Tested in Development)
Mouse BALB/c IgG1, κ
Activated human peripheral blood mononuclear cells
Flow cytometry (Qualified)
0.2 mg/ml
VI 6T-083, M4
AB_2874230
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.
750009 Rev. 4
Antibody Details
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A3

The A3 monoclonal antibody specifically binds to CD148. CD148 is a receptor-type tyrosine phosphatase that belongs to the receptor class 3 subfamily of the protein-tyrosine phosphatase family. CD148 is also known as Density-enhanced phosphatase 1 (DEP-1), Human protein tyrosine phosphatase- η (HPTP-η/HPTP-eta), or Protein tyrosine phosphatase receptor type J (PTPRJ). CD148 is a highly glycosylated type I transmembrane protein that is widely expressed on different cell types including monocytes, granulocytes, dendritic cells, thymocytes, T cells, B cells, NK cells, fibroblasts, and platelets. As a protein tyrosine phosphatase with receptor function, CD148 plays a role in regulating the signaling activities of various phosphorylated cellular proteins including the PDGF Receptor, catenins and Src family kinases.  In the immune system, CD148 plays a role in regulating the activities of different cell types. For example, upon T cell activation, CD148 expression is upregulated by T cells. CD148 can subsequently dephosphorylate LAT and PLC-γ signaling proteins and thereby downregulate T cell signaling responses. Immobilized A3 antibody can reportedly augment the proliferation of anti-CD3 antibody-stimulated peripheral blood T cells in culture.

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.

750009 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
750009 Rev.4
Citations & References
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Development References (4)

  1. Baker JE, Majeti R, Tangye SG, Weiss A. Protein tyrosine phosphatase CD148-mediated inhibition of T-cell receptor signal transduction is associated with reduced LAT and phospholipase Cgamma1 phosphorylation. Mol Cell Biol. 2001; 21(7):2393-2403. (Clone-specific: Flow cytometry). View Reference
  2. Scraven B, Hegen M, Autschbach F, Gaya A, Schwarz C, Meuer SC. CD148 (p260 phosphatase) Workshop Panel report. In: Kishimoto T. Tadamitsu Kishimoto .. et al., ed. Leucocyte typing VI : white cell differentiation antigens : proceedings of the sixth international workshop and conference held in Kobe, Japan, 10-14 November 1996. New York: Garland Pub.; 1997:576-580.
  3. Tangye SG, Phillips JH, Lanier LL, de Vries JE, Aversa G. CD148: a receptor-type protein tyrosine phosphatase involved in the regulation of human T cell activation. J Immunol. 1998; 161(7):9249-9255. (Immunogen: (Co)-stimulation, Flow cytometry, Functional assay). View Reference
  4. Whiteford JR, Xian X, Chaussade C, Vanhaesebroeck B, Nourshargh S, Couchman JR. Syndecan-2 is a novel ligand for the protein tyrosine phosphatase receptor CD148. Mol Cell Biol. 2011; 22(19):3609-3624. (Clone-specific: Flow cytometry, Functional assay, Western blot). View Reference
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750009 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.