Description
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.
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. For optimal and reproducible results, BD Horizon Brilliant™ Stain Buffer should be used anytime BD Horizon Brilliant™ dyes are used in a multicolor flow cytometry panel. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. 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).
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. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
3. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
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. BD Horizon Brilliant Ultraviolet 395 is covered by one or more of the following US patents: 8,158,444; 8,575,303; 8,354,239.
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. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
10. For U.S. patents that may apply, see bd.com/patents.
Specifications

General

SourceMouse TNFSF18, amino acids (Thr47 - Ser173) (Accession# NM_005092), was expressed in 293E cells. The amino-terminal possesses a linker-9His tag.
Molecular MassThe 148 amino acid recombinant protein has a predicted molecular mass of approximately 16.7 kD. The protein migrates at approximately 20-30 kD in both DTT-reducing and non-reducing conditions by SDS-PAGE. The predicted N-terminal amino acid is His.
Purity>95%, as determined by Coomassie stained SDS-PAGE.
Formulation0.22 µm filtered protein solution is in PBS, pH 7.4.
Endotoxin LevelLess than 0.01 ng per µg cytokine as determined by the LAL method.
Concentration10 and 25 µg sizes are bottled at 200 µg/mL. 100 µg size and larger sizes are lot-specific and bottled at the concentration indicated on the vial. To obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.
Storage & HandlingUnopened vial can be stored between 2°C and 8°C for up to 2 weeks, at -20°C for up to six months, or at -70°C or colder until the expiration date. For maximum results, quick spin vial prior to opening. The protein can be aliquoted and stored at -20°C or colder. Stock solutions can also be prepared at 50 - 100 µg/mL in appropriate sterile buffer, carrier protein such as 0.2 - 1% BSA or HSA can be added when preparing the stock solution. Aliquots can be stored between 2°C and 8°C for up to one week and stored at -20°C or colder for up to 3 months. Avoid repeated freeze/thaw cycles.
ActivityED50 = 0.05 - 0.4 µg/mL as determined by the induction of splenocytes proliferation when anti-mouse CD3 (200 ng/well, Cat. No. 100207) and TNFSF18 are immobilized.
ApplicationBioassay
Application NotesBioLegend carrier-free recombinant proteins provided in liquid format are shipped on blue-ice. Our comparison testing data indicates that when handled and stored as recommended, the liquid format has equal or better stability and shelf-life compared to commercially available lyophilized proteins after reconstitution. Our liquid proteins are verified in-house to maintain activity after shipping on blue ice and are backed by our 100% satisfaction guarantee. If you have any concerns, contact us at [email protected].
StructureDimer.
DistributionImmature and mature dendritic cells, B cells, macrophages, keratinocytes, endothelial cells, and retinal epithelial cells.
FunctionThe GITR-GITRL interaction plays an important role in the differentiation of thymic Tregs (tTregs) and the expansion of Tregs, including both tTregs and peripheral Tregs (pTregs). GITRL expression increases during T cell activation and its costimulatory function results in increased TCR-dependent cell proliferation. GITR-GITRL interaction reverses regulatory T cell suppression, which results in greater immunity to tumors and viral pathogens, as well as exacerbating autoimmune diseases.
InteractionActivated CD4+ and CD8+ effector T cells, regulatory T cells, follicular T helper cells, regulatory follicular T helper cells, and natural killer cells.
Ligand/ReceptorTNFRSF18 (GITR, CD357).
Cell TypeEmbryonic Stem Cells
Biology AreaCell Proliferation and Viability, Stem Cells
Molecular FamilyCytokines/Chemokines, Growth Factors
Antigen References1. Nocentini G, et al. 1997. Proc. Natl. Acad. Sci. USA 12:6216. 2. Shimizu J, et al. 2002. Nat. Immunol. 3:135. 3. Yu KY, et al. 2003. Biochem. Biophys. Res. Commun. 310:433. 4. Kim JD, et al. 2003. Genes Immun. 4:564. 5. Kim BJ, et al. 2004. Invest Ophthalmol Vis. Sci. 45:3170. 6. Grohmann U, et al. 2007. Nat. Med. 13:579. 7. Ronchetti S, et al. 2007. J. Immunol. 179:5916. 8. Chattopadhyay K, et al. 2008. Proc. Natl. Acad. Sci. USA 105:635. 9. Byrne AM, et al. 2012. Clin. Exp. Allergy 42:550. 10. Clouthier DL, Watts TH. 2014. Cytokine Growth Factor Rev. 25:91.
Gene ID240873
UniProtView information about TNFSF18 on UniProt.org
Regulatory StatusRUO
Other NamesActivation-inducible TNF receptor (AITR) ligand (AITRL), GITR ligand, GITRL