ULK3 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon1 and 1 bp insertion in exon1.
DKFZP434C131, FLJ90566, Serine/threonine-protein kinase ULK3, ULK3_HUMAN, Unc-51-like kinase 3, unc 51 like kinase 3 (C. elegans)
ULK3 KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon1 and 1 bp insertion in exon1.
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Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.
ULK3 also known as Unc-51 Like Autophagy Activating Kinase 3 is a serine/threonine-protein kinase with a molecular mass of approximately 58 kDa. This protein plays an important role in mediating cellular processes through phosphorylation. ULK3 shows expression in various tissues including the brain and kidney. Its catalytic activity influences multiple cellular events especially those related to autophagy the cell's degradation pathway for removing damaged organelles and recycling cellular components.
This protein contributes significantly through its involvement in autophagy regulation. ULK3 acts as a kinase within a multiprotein complex which includes proteins such as ATG13 and FIP200. This complex is essential for initiating autophagy by regulating the autophagosome formation. Its activity can promote cellular survival under nutrient-deprived conditions showcasing its role as an important regulatory component in cellular maintenance and stress responses.
ULK3 takes part in critical signaling pathways such as the mTOR signaling pathway and the Hedgehog signaling pathway. In the mTOR pathway ULK3 through its kinase activity acts upstream to regulate the activity of the autophagy machinery. It shares a functional relationship with proteins like mTOR and AMPK. In the Hedgehog pathway ULK3 regulates pathway components impacting cellular growth and differentiation processes. This positions ULK3 as a modulator connecting different signaling pathways to maintain cellular homeostasis.
Researchers have reported associations with cancer and neurodegenerative diseases. Abnormal ULK3 activity can influence tumor growth and progression particularly in its regulatory role in cell proliferation and survival. Studies also suggest an involvement in neurodegenerative diseases where dysregulation in autophagy processes in which ULK3 plays a part contributes to neuronal cell death. In these contexts ULK3's interactions with proteins like mTOR and Beclin-1 may exacerbate disease progression by altering normal cellular processes.
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Anti-ULK3 antibody [EPR4888] ab124947 was shown to specifically react with ULK3 in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line Human ULK3 knockout HEK-293T cell line ab266152 (knockout cell lysate ab258271) was used. Wild-type and ULK3 knockout samples were subjected to SDS-PAGE. Anti-ULK3 antibody [EPR4888] ab124947 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4oC at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-ULK3 antibody [EPR4888] (Anti-ULK3 antibody [EPR4888] ab124947) at 1/1000 dilution
Lane 1: Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg
Lane 2: ULK3 knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg
Lane 2: Western blot - Human ULK3 knockout HEK-293T cell line (Human ULK3 knockout HEK-293T cell line ab266152)
Lane 3: HAP1 whole cell lyate at 20 µg
Lane 4: LNCaP (Human prostate cancer cell line) whole cell lysate at 20 µg
All lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/10000 dilution
Predicted band size: 53 kDa
Observed band size: 55 kDa
Allele-2: 1 bp insertion in exon1
Allele-1: 1 bp deletion in exon1
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