Description
The monoclonal antibodies TH6 and 12A5 recognize Folate Receptor 4 (FR4), also known as the membrane folate-binding protein 3 (FBP3). FR4 is a heavily glycosylated 35 kD receptor expressed exclusively in lymphoid tissue and an isoform of the family of receptors that recognize the essential nutrient folic acid. Natural T regulatory cells constitutively express high levels of FR4. Differential expression of FR4 in combination with CD25 can distinguish four functionally distinct CD4+ T cell subpopulations; Natural Tregs, effector T cells, memory-like T cells and Naïve T cells. FR4hi CD25+ expressing CD4+ T cells also express high amounts of Foxp3, GITR and CTLA-4. Monoclonal antibody TH6 and 12A5 stained CD25+CD4+ T cells at a higher level than other CD4+ or CD8+ T cells. In addition, in vivo injection of TH6 monoclonal antibody reduced the number of CD25+CD4+ T cells and CD25-CD4+ T cells in peripheral blood. Clone 12A5 has been demonstrated to work in western blot. Clones TH6 and 12A5 do not block one another in a flow cytometric assay.
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 or 566349). When setting up compensation, it is recommended to compare spillover values obtained from cells and BD™ CompBeads to ensure that beads will provide sufficiently accurate spillover values. 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.
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 BB700 under optimal conditions that minimize unconjugated dye and antibody.
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 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 GE Healthcare.
