Description: The monoclonal antibody A1exF5 recognizes both human and mouse Arginase 1, a cytosolic enzyme (Arg1). This A1exF5 clone is compatible with both, the standard intracellular protocols, and the Foxp3/Transcription Factor Staining Buffer Set. Applications Reported: This A1exF5 antibody has been reported for use in flow cytometric analysis. Applications Tested: This A1exF5 antibody has been tested by flow cytometric analysis of stimulated mouse bone marrow cells using the Intracellular Fixation & Permeabilization Buffer Set (Product No. 88-8824) and protocol. Please refer to Best Protocols: Protocol A: Two step protocol for (cytoplasmic) intracellular proteins located under the Resources Tab online. This may be used at less than or equal to 0.25 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. PerCP-eFluor 710 emits at 710 nm and is excited with the blue laser (488 nm); it can be used in place of PerCP-Cyanine5.5. We recommend using a 710/50 bandpass filter, however, the 695/40 bandpass filter is an acceptable alternative. Please make sure that your instrument is capable of detecting this fluorochrome.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 (Product No. 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product No. 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 nm; Emission: 710 nm; Laser: Blue Laser Arginase-1 (Arg1) is a 35 kDa enzyme converting L-arginine to urea and L-ornithine, which is the final step in the urea cycle. The resulting polyamines are important for cell proliferation and removal of toxins that arise from protein degradation. By degrading arginine, Arginase 1 deprives NO synthase of its substrate and down-regulates nitric oxide production. In both human and mouse, Arginase 1 is expressed in the liver, neutrophils, myeloid derived suppressor cells (MDSC) and neural stem cells. In human, expression in blood neutrophils but not in CCR3+ granulocytes has been reported. In mice, expression of Arginase 1 is one of the hallmarks of alternatively activated macrophages (M2a). Arginase-1 may be expressed in the myeloid cells infiltrating tumors, and is typically found in the majority of hepatocellular carcinomas. Defects in Arginase 1 are the cause of argininemia, an autosomal recessive disorder characterized by hyperammonemia.
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