c-MAF Monoclonal Antibody (sym0F1), PE-eFluor™ 610, eBioscience™, Invitrogen™

Description

Description: The sym0F1 monoclonal antibody reacts with human and mouse c-Maf, a 42 kDa basic leucine zipper transcription factor shown to be involved in the neural, ocular and hematopoietic systems. In hematopoietic cells, it was first shown to be crucial for IL-4 expression in Th2 and was the first transcription factor believed to be Th subset-specific. More recent evidence shows that specific phospho-tyrosine residues lead to upregulation of IL-4. c-Maf has also been shown to be important to differentiation and function in both Th17 and Tfh cells. It drives expression of IL-21 in both cell types, while promoting expression of IL-23R in Th17 and CXCR5 in Tfh as well. Sym0F1 does not cross-react with MafA or MafB. Applications Reported: This sym0F1 antibody has been reported for use in intracellular staining followed by flow cytometric analysis. Applications Tested: This sym0F1 antibody has been pre-diluted and tested by intracellular staining followed by flow cytometric analysis of Th17-polarized mouse splenocytes using the Foxp3/Transcription Factor Staining Buffer Set (Product # 00-5523-00) and protocol. Please refer to 'Staining Intracellular Antigens for Flow Cytometry, Protocol B: One step protocol for intracellular (nuclear) proteins' located at Flow Protocols. This may be used at 5 μL (1.0 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. PE-eFluor 610 can be excited with laser lines from 488-561 nm and emits at 607 nm. We recommend using a 610/20 band pass filter (equivalent to PE-Texas Red). 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 # 00-8222-49) (100 μL of cell sample + 100 μL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333-57) 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: 607 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser

c-maf is the cellular counterpart of oncogenic v-maf that belongs to the family of basic region leucine zipper domain transcription factors. The leucine-zipper domain is involved in the interaction with LRPICD. There are two forms of human c-maf mRNA, c-maf-long and c-maf-short. It is identified in the genome of the acute transforming avian retrovirus AS42. c-maf targets are IL-4 in Th2 cells, the crystalline genes in lens fiber cells, insulin gene in islet , p53 and L7 where it exerts its transcriptional role through binding to a Maf recognition element (MARE). It regulates Th2 differentiation and lineage-specific hematopoiesis. c-maf is a transcription factor for IL-10 gene expression in LPS-activated macrophages. Chromosomal aberration involving maf is found in some forms of multiple myeloma. It is expressed in myeloma cell lines and resting monocytes/macrophages.