-
Your selected country is
Portugal
- Change country/language
Old Browser
This page has been recently translated and is available in French now.
Looks like you're visiting us from {countryName}.
Would you like to stay on the current country site or be switched to your country?
Two-color flow cytometric analysis of CD273 (PD-L2) expression on mouse splenic dendritic cells. BALB/c mouse splenic leucocytes were cultured overnight with Recombinant Mouse GM-CSF protein (Cat. No. 554586; 5 ng/ml). The cells were harvested and preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with APC Hamster Anti-Mouse CD11c antibody (Cat. No. 550261/561119) and either PE Rat IgG2a, κ Isotype Control (Cat. No. 553930; Left Plot) or PE Rat Anti-Mouse CD273 (Cat. No. 567217; Right Plot) at 0.25μg/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 CD273 (PD-L2) [or Ig Isotype control staining] versus CD11c was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) splenic leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
BD Pharmingen™ PE Rat Anti-Mouse CD273 (PD-L2)
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
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 cell and CompBead to ensure that BD Comp beads are appropriate for your specific cellular application.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
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.
Development References (6)
-
Loke P, Allison JP. PD-L1 and PD-L2 are differentially regulated by Th1 and Th2 cells.. Proc Natl Acad Sci U S A. 2003; 100(9):5336-41. (Biology). View Reference
-
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: In vivo exacerbation). View Reference
-
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: Functional assay, In vivo exacerbation). View Reference
-
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, Functional assay, Inhibition, In vivo exacerbation). View Reference
-
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: Blocking). View Reference
-
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). View Reference
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.