|Detection of Human, Mouse, and Rat UFM1 by Western Blot. Western blot shows lysates of HepG2 human hepatocellular carcinoma cell line, C2C12 mouse myoblast cell line, and Rat‑2 rat embryonic fibroblast cell line. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human UFM1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8237) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for UFM1 at approximately 10 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.|
|UFM1 in HeLa Human Cell Line. UFM1 was detected in immersion fixed HeLa human cervical epithelial carcinoma cell line using Sheep Anti-Human UFM1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF8237) at 1.7 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red, upper panel; Catalog # NL010) and counterstained with DAPI (blue, lower panel). Specific staining was localized to nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.|
UFM1 ( Ubiquitin-fold modifier 1) is a 9.1 kDa ubiquitin-like protein, displaying a similar tertiary structure to ubiquitin. The UFM1 conjugation system is a novel Ubiquitin-like (Ubl) system whose physiological target(s) and biological functions remain largely undefined. To be activated, UFM1 is processed C-terminally by two specific proteases, UfSP1 and UfSP2. After processing, UFM1 is activated via the E1 enzyme, UBA5, and then conjugated by the E2 enzyme, UFC1. UFL1 has been identified as the E3 enzyme. However, cellular functions associated with target proteins that are modified by UFM1 are still unknown. Genetic study has demonstrated that the Ufm1-activating enzyme Uba5 is indispensible for erythroid differentiation in mice, highlighting the importance of this novel system in animal development. It was found that the UFM1 system was transcriptionally up-regulated by disturbance of the ER homeostasis and inhibition of vesicle trafficking.
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