Rabbit Recombinant Monoclonal PICALM antibody. Carrier free. Suitable for WB, IHC-P, ICC/IF and reacts with Human samples.
pH: 7.2 - 7.4
Constituents: PBS
IP | Flow Cyt | WB | IHC-P | ICC/IF | |
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Human | Not recommended | Not recommended | Tested | Expected | Expected |
Mouse | Not recommended | Not recommended | Predicted | Predicted | Predicted |
Species | Dilution info | Notes |
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Species Human, Mouse | Dilution info - | Notes - |
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Species Human, Mouse | Dilution info - | Notes - |
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Species Human | Dilution info - | Notes - |
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Species Mouse | Dilution info - | Notes - |
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Species Human | Dilution info - | Notes Perform heat-mediated antigen retrieval before commencing with IHC staining protocol. |
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Species Mouse | Dilution info - | Notes - |
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Species Human | Dilution info Use at an assay dependent concentration. | Notes - |
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Species Mouse | Dilution info - | Notes - |
Cytoplasmic adapter protein that plays a critical role in clathrin-mediated endocytosis which is important in processes such as internalization of cell receptors, synaptic transmission or removal of apoptotic cells. Recruits AP-2 and attaches clathrin triskelions to the cytoplasmic side of plasma membrane leading to clathrin-coated vesicles (CCVs) assembly (PubMed:10436022, PubMed:16262731, PubMed:27574975). Furthermore, regulates clathrin-coated vesicle size and maturation by directly sensing and driving membrane curvature (PubMed:25898166). In addition to binding to clathrin, mediates the endocytosis of small R-SNARES (Soluble NSF Attachment Protein REceptors) between plasma membranes and endosomes including VAMP2, VAMP3, VAMP4, VAMP7 or VAMP8 (PubMed:21808019, PubMed:22118466, PubMed:23741335). In turn, PICALM-dependent SNARE endocytosis is required for the formation and maturation of autophagic precursors (PubMed:25241929). Modulates thereby autophagy and the turnover of autophagy substrates such as MAPT/TAU or amyloid precursor protein cleaved C-terminal fragment (APP-CTF) (PubMed:24067654, PubMed:25241929).
CALM, PICALM, Phosphatidylinositol-binding clathrin assembly protein, Clathrin assembly lymphoid myeloid leukemia protein
Rabbit Recombinant Monoclonal PICALM antibody. Carrier free. Suitable for WB, IHC-P, ICC/IF and reacts with Human samples.
pH: 7.2 - 7.4
Constituents: PBS
ab249736 is the carrier-free version of Anti-PICALM antibody [EPR12177] ab172962.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
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This data was developed using the same antibody clone in a different buffer formulation (Anti-PICALM antibody [EPR12177] ab172962).
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 Human PICALM knockout HeLa 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
This data was developed using the same antibody clone in a different buffer formulation (Anti-PICALM antibody [EPR12177] ab172962).
Lanes 1 - 4: Merged signal (red and green). Green - Anti-PICALM antibody [EPR12177] ab172962 observed at 70-75 kDa. Red - loading control, Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.
Anti-PICALM antibody [EPR12177] ab172962 was shown to react with PICALM in U-2 OS wild-type cells in Western blot. Loss of signal was observed when PICALM knockout sample was used. U-2 OS wild-type and PICALM knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% Milk in TBS-T (0.1% Tween®) before incubation with Anti-PICALM antibody [EPR12177] ab172962 and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4°C at a 1 in 5000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (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 1/5000 dilution
Lane 1: Wild-type U-2 OS whole cell lysate at 20 µg
Lane 2: PICALM knockout U-2 OS whole cell lysate at 20 µg
Lane 2: Western blot - Human PICALM knockout U-2 OS cell line (Human PICALM knockout U-2 OS cell line ab262500)
Lane 3: PC3 (Human prostate adenocarcinoma cell line) whole cell lysate at 20 µg
Lane 4: Caco-2 (Human colorectal adenocarcinoma cell line) whole cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 70 kDa
Observed band size: 70-75 kDa
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