• Product name

  • Description

    Rabbit polyclonal to Piccolo
  • Host species

  • Specificity

    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.
  • Tested applications

    Suitable for: IHC-Fr, ICC, IHC (PFA fixed), WB, IHC-FoFr, IHC-Pmore details
  • Species reactivity

    Reacts with: Rat
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 600 - 700 of Rat Piccolo.

    Read Abcam's proprietary immunogen policy (Peptide available as ab23400.)

  • Positive control

    • ab20664 gave a positive result in the following tissue lysates: Rat brain and cerebellum.


  • Form

  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

  • Isotype

  • Research areas


Our Abpromise guarantee covers the use of ab20664 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-Fr 1/500. PubMed: 19218615
ICC Use at an assay dependent concentration. PubMed: 19377471
IHC (PFA fixed) 1/300 - 1/1000.
WB Use a concentration of 0.2 - 0.5 µg/ml. Detects a band of approximately 460 kDa (predicted molecular weight: 520 kDa).

Abcam recommends using milk as the blocking agent.

IHC-FoFr 1/300.
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.


  • Function

    May act as a scaffolding protein involved in the organization of synaptic active zones and in synaptic vesicle trafficking.
  • Sequence similarities

    Contains 2 C2 domains.
    Contains 1 PDZ (DHR) domain.
  • Domain

    C2 domain 1 is involved in binding calcium and phospholipids. Calcium binds with low affinity but with high specificity and induces a large conformational change.
  • Cellular localization

    Cell junction > synapse. Concentrated at the presynaptic side of synaptic junctions.
  • Information by UniProt
  • Database links

  • Alternative names

    • ACZ antibody
    • Aczonin antibody
    • Brain-derived HLMN protein antibody
    • DKFZp779G1236 antibody
    • KIAA0559 antibody
    • Multidomain presynaptic cytomatrix protein antibody
    • PCLO antibody
    • PCLO_HUMAN antibody
    • Piccolo (presynaptic cytomatrix protein) antibody
    • Piccolo, mouse, homolog of antibody
    • Pico antibody
    • Presynaptic cytomatrix protein antibody
    • Protein piccolo antibody
    see all


  • IHC image of Piccolo staining in rat brain FFPE section, performed on a BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab20664, 5µg/ml, for 8 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
  • All lanes : Anti-Piccolo antibody (ab20664) at 1 µg/ml

    Lane 1 : Brain (Rat) Tissue Lysate
    Lane 2 : Cerebellum (Rat) Tissue Lysate
    Lane 3 : Kidney (Rat) Tissue Lysate (Negative control)

    Lysates/proteins at 10 µg per lane.

    All lanes : Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/50000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 520 kDa
    Observed band size: 460 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 200 kDa. We are unsure as to the identity of these extra bands.

    Exposure time: 4 minutes

    The banding pattern shown in the image above is consistent with the literature which describes multiple bands >420 kDa as a result of proteolysis (PMID : 10707984).

    This blot was produced using a 3-8% Tris Acetate gel under the TA buffer system. The gel was run at 150V for 60 minutes using lysates heated to 85°C before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% Milk before being incubated with ab20664 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

  • IHC-FoFr image of Piccolo staining on rat brain cortex tissue sections. The tissues were fixed (animals perfused fixed) with 4% PFA and later postfixed overnight in the same fixative. They were cryoprotected in 30% sucrose and cut using a cryostat. The staining was performed using free floating technique.

    See Abreview

  • Immuofluorescent staining for Piccolo ab20664 in [A] rat brain hippocampus (X20 objective) and [B] rat brain cortex (X40 objective). Tissue preparation: rat brain tissue was perfusion fixed (4% PFA) followed by post fix and cryoprotection in 20% sucrose before freezing in OCT. 30µm coronal sections were cut on a cryostat for free floating IHC. Primary antibody ab20664 was used at 1/100 (5µg/ml) incubated overnight at room temperature in PBST (triton 0.3%). Secondary antibody used: anti-rabbit Alexa fluor 488 (1/1000) incubated for 2 hours at room temperature.
  • All lanes : Anti-Piccolo antibody (ab20664) at 0.2 µg/ml

    Lane 1 : Rat brain lysate at 40 µg
    Lane 2 : Mouse brain lysate at 50 µg

    All lanes : 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.


This product has been referenced in:

See all 6 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Human Tissue sections (Brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: PerkinElmer AR6
Blocking step
Serum as blocking agent for 45 minute(s) · Concentration: 10% · Temperature: 20°C

Dr. An Truong

Verified customer

Submitted Jun 19 2019

Immunohistochemistry (PFA perfusion fixed frozen sections)
Rat Tissue sections (Brain)

Dr. Karine Thibault

Verified customer

Submitted Dec 19 2012


Thank you for contacting us.

I am not sure how PSD95 expression would be affected if toxicity is increased in your cells would not wantto recommend a product that may be unsuitable. We encourage customers to consult the latest literature available through PubMed and other resources in order to find the most up-to-date information about their specific research interests.

I am sorry that I could not be more helpful, but I hope that the available literature in this area can provide some clarification. Please do not hesitate to contact us again with other needs or with any questions about our products.

Read More


Thank you for contacting Abcam.

In mouse neurons an alternative to the smaller housekeeping proteins such as beta-actin, or GAPDH commonly used as loading control would be to use synaptic markers such as PSD95 or Piccolo.

In western blot I would recommend use of anti-PSD95 antibodieshttps://www.abcam.com/PSD95-antibody-Synaptic-Marker-ab18258.htmlorhttps://www.abcam.com/PSD95-antibody-6G6-1C9-ab2723.htmlwhich will give you band above your threshold size at 95kDA.

Should you want bands with an even higher molecular weight Piccolo has a predicted size at 540kDA, our producthttps://www.abcam.com/Piccolo-antibody-Synaptic-Marker-ab20664.htmlis, like the above PSD95 antibodies, guaranteed to work on mouse samples in Western blot.

I hope that this information is helpful. Please let me know if you have any questions or there are other ways that Abcam may help you meet your research goals.

Use our products? Submit an Abreview. Earn rewards!


Read More
Immunohistochemistry (Frozen sections)
Rat Tissue sections (Sections of rat brain)
Sections of rat brain

Dr. Sophie Pezet

Verified customer

Submitted May 18 2006

Western blot
Rat Tissue lysate - whole (Whole brain lysate - adult)
Loading amount
40 µg
Whole brain lysate - adult
Blocking step
Milk as blocking agent for 20 minute(s) · Concentration: 5%

Dr. Randal Moldrich

Verified customer

Submitted May 18 2006

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