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Western blot - Piccolo antibody - Synaptic Marker (ab20664)
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Piccolo antibody - Synaptic Marker (ab20664)
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Immunohistochemistry (PFA fixed) - Piccolo antibody - Synaptic Marker (ab20664)
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Product Name
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Piccolo antibody - Synaptic Marker
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See all Piccolo antibodies (3)...
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Product type
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Primary antibodies
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Description
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Rabbit polyclonal to Piccolo - Synaptic Marker
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Immunogen
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Synthetic peptide conjugated to KLH derived from within residues 600 - 700 of Rat Piccolo.
(Peptide available as ab23400.)
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Reacts with
(species key)
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Ms, Rat
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Specificity
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Due to its large size, Piccolo requires special gel-electrophoresis and Western blotting protocols for visualization by immunoblotting.
Excellent results can be obtained, for example, with the 4-12% TRIS-glycine gradient gels of Anamed. For success in WB with Rabbit polyclonal to Piccolo - Synaptic Marker (ab20664), do not denature WB sample lysate.
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Tested applications
(see key)
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ICC, IHC (PFA fixed), IHC-Fr, IHC-P, WB
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Abreviews
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Application notes
(see key)
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Recommended dilutions
ICC: Use at an assay dependent dilution (PMID 19377471).
IHC-P: Use at a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IHC-Fr: 1/500 (PMID 19218615).
Immunohistochemistry (PFA fixed): 1/300 - 1/1000.
WB: Use at a concentration of 0.2-0.5 µg/ml. Detects a band of approximately 520 kDa (predicted molecular weight: 520 kDa).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
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Positive control
(see definition)
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Rat brain and mouse brain
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Cellular localization
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Cytoplasmic
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Research areas
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Neuroscience >> Cell Type Marker >> Neuron marker >> Synapse marker
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Relevance
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Piccolo is a presynaptic cytomatrix protein concentrated at the presynaptic side of synaptic junctions. Piccolo is a large protein which constists of an N-terminal Zn2+ finger, several piccolo-bassoon homology domains (PBH-domains) and C-terminal PDZ and C2 domains. It is usually found together with bassoon, a related huge multi-domain protein of the CAZ (cytoskeletal matric assembled at active zones). Piccolo is a scaffolding protein for proteins involved in endo- and exocytosis of synaptic vesicles. Recently piccolo has been shown to interfere with clathrin mediated endocytosis by binding to the F-actin and dynamin binding protein Abp1. Piccolo is highly expressed in brain with low levels found in stomach. It is not detected in other tissues analyzed including adrenal gland, testis and pancreas.
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Database links
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The links below go to external sites and will open in a new browser window |
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Raised in
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Rabbit
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Clonality
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Polyclonal
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Isotype
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IgG
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Purity
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Immunogen affinity purified
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Storage buffer
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Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 Material safety datasheet (MSDS) for this product:
Sodium Azide MSDS
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Form
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Liquid
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Concentration
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0.360 mg/ml
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Storage instructions
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Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
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At Abcam, we have one centralized database to hold all of our product information, so that everything we know about this Piccolo antibody - Synaptic Marker is on this datasheet. But please do contact us if you would like any reassurance! |
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See below for Piccolo antibody - Synaptic Marker images, references, products related to ab20664 and other tools.
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Piccolo antibody - Synaptic Marker images:
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 Western blot - Piccolo antibody - Synaptic Marker (ab20664)
All lanes : Piccolo antibody - Synaptic Marker (ab20664) at 0.2 µg/ml
Lane 1 : Rat brain lysate at 40 µg
Lane 2 : Mouse brain lysate at 50 µg
Secondary
Anti-rabbit HRP at 1/20000 dilution
developed using the ECL technique
Performed under reducing conditions.
Predicted band size : 520 kDa
Observed band size : 520 kDa
Additional bands at : 60 kDa. We are unsure as to the identity of these extra bands.
Exposure time : 10 minutes
Primary Antibody : anti-Piccolo (ab20664) for 1h in 5% Whole Milk Powder (WMP); Secondary Antibody : Anti-rabbit HRP (1/20000) for 1h in 5% WMP; Migration medium: Laemmeli + glycerol + 5% B-mercaptoethanol; Transfer : Tris/Glycine, 20% ethanol NB: Gels higher than 8% acrylamide were tried without success; 8% or lower is recommended. Denaturing of samples at 70C or 95C was not successful. Reducing conditions were used and 20% ethanol was employed for the nitrocellulose membrane transfer. All steps performed at RT. | |
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Pricing information loading
This product has been used in: (two most recent references)