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PE Rat Anti-Mouse MD-1
PE Rat Anti-Mouse MD-1

Multicolor flow cytometric analysis of MD-1 expression on viable Mouse splenic leucocytes.  C57BL/6 Mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142]. The splenic leucocytes were then stained with APC Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 553092) and with either PE Rat IgG2b, κ Isotype Control (Cat. No. 555848; Left Plot) or PE Rat Anti-Mouse MD-1 antibody (Cat. No. 569972/569973; Right Plot) at 0.25 μg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of MD-1 versus CD45R/B220 (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) cells. Flow cytometric analysis was performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

Multicolor flow cytometric analysis of MD-1 expression on viable Mouse splenic leucocytes.  C57BL/6 Mouse splenic leucocytes were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142]. The splenic leucocytes were then stained with APC Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 553092) and with either PE Rat IgG2b, κ Isotype Control (Cat. No. 555848; Left Plot) or PE Rat Anti-Mouse MD-1 antibody (Cat. No. 569972/569973; Right Plot) at 0.25 μg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of MD-1 versus CD45R/B220 (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) cells. Flow cytometric analysis was 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 Pharmingen™
Ly86; MD-1; MD1; ly-86; lymphocyte antigen 86
Mouse (QC Testing)
Rat IgG2b, κ
Mouse RP105/MD-1 Transfected Cells
Flow cytometry (Routinely Tested)
0.2 mg/ml
17084
AB_3685445
Aqueous buffered solution containing protein stabilizer 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 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 cell 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. 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.
  3. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  5. An isotype control should be used at the same concentration as the antibody of interest.
  6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  7. For U.S. patents that may apply, see bd.com/patents.
569973 Rev. 1
Antibody Details
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MD113

The MD113 monoclonal antibody specifically recognizes MD-1, which is also known as Ly-86. MD-1 is encoded by Ly86 (lymphocyte antigen 86). MD-1 is expressed by B cells, dendritic cells (DCs), monocytes, and macrophages. MD-1 is an ~22-25 kDa secreted glycoprotein that complexes with the extracellular domain of the transmembrane Radioprotective 105 kDa protein (RP105) also known as CD180. In combination with TLR4 (CD284), the MD-1/RP105 complex mediates the B cell response to stimulation by lipopolysaccharide (LPS) that results in the upregulated expression of the B7.2 (CD86) costimulatory molecule, cytokine production, proliferation, and resistance to apoptosis.

569973 Rev. 1
Format Details
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
PE
Yellow-Green 561 nm
496 nm, 566 nm
576 nm
569973 Rev.1
Citations & References
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View product citations for antibody "569973" on CiteAb

Development References (3)

  1. Miyake K, Shimazu R, Kondo J, et al. Mouse MD-1, a molecule that is physically associated with RP105 and positively regulates its expression.. J Immunol. 1998; 161(3):1348-1353. (Biology). View Reference
  2. Miyake K, Yamashita Y, Ogata M, Sudo T, Kimoto M. RP105, a novel B cell surface molecule implicated in B cell activation, is a member of the leucine-rich repeat protein family. J Immunol. 1995; 154(7):3333-3340. (Biology). View Reference
  3. Nagai Y, Shimazu R, Ogata H et al. Requirement for MD-1 in cell surface expression of RP105/CD180 and B-cell responsiveness to lipopolysaccharide. Blood. 2002; 99(5):1699-1705. (Immunogen: Functional assay, Inhibition). View Reference
569973 Rev. 1

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