PICALM 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 exon4 and 5 bp deletion in exon4.
CLTH, Clathrin assembly lymphoid myeloid leukemia, Clathrin assembly lymphoid myeloid leukemia protein, LAP, PICAL_HUMAN, Phosphatidylinositol-binding clathrin assembly protein
PICALM 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 exon4 and 5 bp deletion in exon4.
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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.
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PICALM also known as Phosphatidylinositol binding clathrin assembly protein is a protein weighing approximately 70 kDa. It plays an important role in clathrin-mediated endocytosis where it helps assembly of the clathrin coat on the cytoplasmic face of the cell membrane. Expression of the PICALM gene is found in various tissues with significant levels in the brain lungs and spleen. The gene encoding PICALM is located on chromosome 11.
PICALM is essential in the trafficking of molecules within cells particularly in the endocytosis and endosomal sorting pathways. This protein is part of large protein complexes assembled during the formation of vesicles. Through these complexes PICALM influences the transport and recycling of membrane receptors and other molecules ensuring proper cellular functions. The clathrin coat assembly and disassembly rely heavily on its function for effective cellular trafficking processes.
PICALM integrates into the clathrin-mediated endocytosis and vesicle-mediated transport pathways. It works alongside proteins such as adaptin and dynamin to facilitate the internalization of proteins and later processing within cellular compartments. In the context of cellular signaling and homeostasis these pathways involving PICALM contribute significantly to receptor recycling and downregulation maintaining proper cellular responses to external stimuli.
PICALM has been linked to Alzheimer's disease and acute myeloid leukemia (AML). In Alzheimer's disease altered PICALM function or expression can affect amyloid precursor protein processing implicating proteins such as presenilins and tau as well. For acute myeloid leukemia the PICALM-MLLT10 fusion protein resulting from chromosomal translocations can contribute to leukemogenesis by disrupting normal gene regulation. These associations highlight the diverse roles and importance of PICALM in both normal physiology and pathology.
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Lane 2: PICALM knockout HeLa cell lysate (20 μg)
Lane 3: Caco-2 cell lysate (20 μg)
Lanes 1-3: Merged signal (red and green). Green - Anti-PICALM antibody [EPR12177] ab172962 observed at 70-75 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 36 kDa.
Anti-PICALM antibody [EPR12177] ab172962 Anti-PICALM antibody [EPR12177] was shown to specifically react with PICALM in wild-type HeLa cells. Loss of signal was observed when knockout cell line Human PICALM knockout HeLa cell line ab265653 (knockout cell lysate ab258113) was used. Wild-type and PICALM knockout samples were subjected to SDS-PAGE. Anti-PICALM antibody [EPR12177] ab172962 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4°C 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-PICALM antibody [EPR12177] (Anti-PICALM antibody [EPR12177] ab172962) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: PICALM knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human PICALM knockout HeLa cell line (Human PICALM knockout HeLa cell line ab265653)
Lane 3: Caco-2 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: 70 kDa
Observed band size: 70-75 kDa
Allele-2: 1 bp deletion in exon4
Allele-1: 5 bp deletion in exon4
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