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BB700 Rat Anti-Mouse CD273 (PD-L2)
Product Details
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BD OptiBuild™
Mouse (Tested in Development)
Rat SD, also known as Sprague-Dawley (outbred) IgG2a, κ
Mouse B7-DC-transfected Cell Line
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 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 CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.

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. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

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. 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.
  2. Researchers should determine the optimal concentration of this reagent for their individual applications.
  3. An isotype control should be used at the same concentration as the antibody of interest.
  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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  7. 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.
  8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  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 Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
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Antibody Details
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MIH37

The MIH37 monoclonal antibody specifically recognizes Programmed cell death 1 ligand 2 (PD-L2) which is also known as CD273 and Butyrophilin B7-DC (B7-DC). This regulatory molecule is a 42-kDa type I membrane glycoprotein encoded by Pdcd1lg2 which belongs to the B7 family of the Ig superfamily. CD273 (PD-L2)  is comprised of an extracellular region with an N-terminal IgV-like domain followed by an IgC2-type domain, a transmembrane sequence, and a cytoplasmic tail. Although not detected on resting leucocytes, its expression is upregulated upon activation of macrophages and dendritic cells (DC) by a variety of stimulatory factors including IL-4, IL-13, or GM-CSF. CD273 (PD-L2) serves as a ligand for the coinhibitory receptor, CD279 (PD-1), a receptor that likewise binds to the CD274 (PD-L1) ligand. CD273 (PD-L2) also binds to Repulsive guidance molecule b (RGMb) that is expressed by T cells, macrophages, neutrophils, and DC. RGMb may associate with other receptors that transduce costimulatory or coinhibitory signals. The MIH37 antibody reportedly blocks the binding of CD273 (PB-L2) to its receptors, CD279 (PD-1) or RGMb and can inhibit antigen-specific T cell responses and hapten-induced contact hypersensitivity reactions. The TY25 monoclonal antibody reportedly binds with lower affinity near or at the same mouse CD273 (PD-L2) epitope recognized by the MIH37 antibody.

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 (6)

  1. Matsumoto K, Fukuyama S, Eguchi-Tsuda M, et al. B7-DC induced by IL-13 works as a feedback regulator in the effector phase of allergic asthma.. Biochem Biophys Res Commun. 2008; 365(1):170-5. (Clone-specific: Flow cytometry). View Reference
  2. Nie X, Chen W, Zhu Y, et al. B7-DC (PD-L2) costimulation of CD4(+) T-helper 1 response via RGMb. Cell Mol Immunol. 2017; 14:1-10. (Biology). View Reference
  3. Okudaira K, Hokari R, Tsuzuki Y, et al. Blockade of B7-H1 or B7-DC induces an anti-tumor effect in a mouse pancreatic cancer model. Int J Oncol. 2009; 35(4):741-749. (Clone-specific: Flow cytometry). View Reference
  4. Ritprajak P, Hashiguchi M, Akiba H, Yagita H, Okumura K, Azuma M. Antibodies against B7-DC with differential binding properties exert opposite effects. Hybridoma. 2012; 31(1):40-47. (Immunogen: Flow cytometry). View Reference
  5. Xiao Y, Yu S, Zhu B, et al. RGMb is a novel binding partner for PD-L2 and its engagement with PD-L2 promotes respiratory tolerance. J Exp Med. 2014; 211(5):943-959. (Clone-specific: Flow cytometry). View Reference
  6. Yamazaki T, Akiba H, Iwai H, et al. Expression of programmed death 1 ligands by murine T cells and APC. J Immunol. 2002; 169(10):5538-5545. (Biology: Flow cytometry). View Reference
View All (6) View Less
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Please refer to Support Documents for Quality Certificates


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.

Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product 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.