Description: The monoclonal antibody eBio12A5 recognizes FR4, also known as Folate receptor 4, FR delta and FBP3 (folate binding protein3). FR4 is a heavily glycosylated, 35 kD receptor for folic acid and the physiologic circulating form of the vitamin, N5-methyltetrahydrofolate. Natural Treg have high levels of FR4 and together with CD25 can be used to distinguish natural Treg, effector T cells, memory-like T cells and naive T cells. FR4 high CD25 high cells are natural Treg with high levels of Foxp3. FR4 high CD25 low cells are identified as central memory T cells, which upon stimulation, can proliferate and produce large amounts of IL-2. In contrast, FR4 low CD25 high cells secrete proinflammatory cytokines such as IFN gamma and IL-17 and have been characterized as effector memory T cells. Based on co-staining studies, the epitopes recognized by eBioTH6 and eBio12A5 are different, thereby allowing functional studies with eBioTH6 to be evaluated with eBio12A5. Applications Reported: This eBio12A5 antibody has been reported for use in flow cytometric analysis. Applications Tested: This eBio12A5 antibody has been tested by flow cytyometric analysis of mouse splenocytes. This can be used at less than or equal to 0.5 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light. Fixation: Samples can be stored in IC Fixation Buffer (cat. 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (cat. 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically. Excitation: 488-561 nm; Emission: 775 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser. Filtration: 0.2 µm post-manufacturing filtered. FR4 is a heavily glycosylated 35 kD receptor for folic acid and the physiologic circulating form of the vitamin, N5-methyltetrahydrofolate. Natural T regs have high levels of FR4 and together with CD25 can be used to distinguish natural Tregs, effector T cells, memory-like T cells and naive T cells. In vivo studies with eBioTH6 show a decrease in the number of CD4+CD25+ cells and an enhanced tumor response leading to the elimination of advanced stages of tumors.
|4° C, store in dark, DO NOT FREEZE!|
|PBS with 0.1% gelatin and 0.09% sodium azide; pH 7.2|
For Research Use Only.
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