Anti-Synapsin I antibody - Synaptic Marker (ab64581)

Overview

  • Product nameAnti-Synapsin I antibody - Synaptic Marker
    See all Synapsin I primary antibodies
  • Description
    Rabbit polyclonal to Synapsin I - Synaptic Marker
  • Tested applicationsSuitable for: WB, IHC-P, IHC-FoFr, ICC/IF, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Sheep, Goat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Rat Synapsin I aa 600-700 conjugated to Keyhole Limpet Haemocyanin (KLH). The exact sequence is proprietary.
    (Peptide available as ab64580)

  • Positive control
    • WB: Mouse spinal cord, mouse brain and rat brain tissue lysates. IP: Mouse brain tissue lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab64581 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
WB Use a concentration of 1 µg/ml. Detects a band of approximately 74, 70 kDa (predicted molecular weight: 74 kDa).

Abcam recommends using 3-5% milk as the blocking agent.

IHC-P Use at an assay dependent concentration.
IHC-FoFr 1/100.
ICC/IF 1/200.
IP Use at an assay dependent concentration.

Target

  • FunctionNeuronal phosphoprotein that coats synaptic vesicles, binds to the cytoskeleton, and is believed to function in the regulation of neurotransmitter release. The complex formed with NOS1 and CAPON proteins is necessary for specific nitric-oxid functions at a presynaptic level.
  • Involvement in diseaseDefects in SYN1 are a cause of epilepsy X-linked with variable learning disabilities and behavior disorders [MIM:300491]. XELBD is characterized by variable combinations of epilepsy, learning difficulties, macrocephaly, and aggressive behavior.
  • Sequence similaritiesBelongs to the synapsin family.
  • Post-translational
    modifications
    Substrate of at least four different protein kinases. It is probable that phosphorylation plays a role in the regulation of synapsin-1 in the nerve terminal. Phosphorylated upon DNA damage, probably by ATM or ATR.
  • Cellular localizationCell junction > synapse. Golgi apparatus.
  • Information by UniProt
  • Database links
  • Alternative names
    • Brain protein 4.1 antibody
    • SYN 1 antibody
    • SYN 1a antibody
    • SYN 1b antibody
    • SYN I antibody
    • SYN1 antibody
    • SYN1_HUMAN antibody
    • SYN1a antibody
    • SYN1b antibody
    • Synapsin 1 antibody
    • Synapsin I antibody
    • Synapsin-1 antibody
    • Synapsin1 antibody
    • SynapsinI antibody
    • SYNI antibody
    see all

Anti-Synapsin I antibody - Synaptic Marker images

  • IHC-P image of ab64581 staining of Synapsin 1 in lamb cerebellum. Heat mediated antigen retrieval buffer was performed using citric acid buffer. The sections were then incubated in 1% BSA to block non-specific protein-protein interactions. The sections were incubated with ab64581 (1:100) for two hours at +21°C, followed by biotin conjugated secondary antibody. There appears to be appropriate, identical positivity in lamb, goat, mouse and rat brain.

    See Abreview

  • Anti-Synapsin I antibody - Synaptic Marker (ab64581) at 1/5000 dilution + Rat whole brain tissue lysate at 15 µg

    Secondary
    Donkey anti-Rabbit IgG at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 74 kDa
    Observed band size : 80,85 kDa (why is the actual band size different from the predicted?)


    Exposure time : 15 seconds

    This image is courtesy of an anonymous Abreview

    Blocked with 5% milk for 1 hour at 25°C.

    Sample was incubated with primary antibody for 18 hours at 4°C in TBS + 0.1% Tween.

    See Abreview

  • ab64581 staining cultured rat hippocampal neurons by ICC/IF. The cultured neurons were fixed with methanol, permeabilized with 0.3% Triton X in 0.1% PBS and blocked with 10% donkey serum for 30 minutes at 24°C. The cultured neurons were then stained with ab64581 at 1/200 in 0.3% TritonX with 0.1% PBS and 10% donkey serum for 4h at 24°C. An Alexa Fluro 488 donkey anti-rabbit polyclonal antibody at 1/1000 was used as the secondary antibody. Nuclei were stained with Hoechst and can be observed in blue.

    See Abreview

  • All lanes : Anti-Synapsin I antibody - Synaptic Marker (ab64581) at 1 µg/ml

    Lane 1 : Spinal Cord (Mouse) Tissue Lysate
    Lane 2 : Brain (Mouse) Tissue Lysate
    Lane 3 : Brain (Rat) Tissue Lysate - normal tissue

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size : 74 kDa
    Observed band size : 70,74 kDa (why is the actual band size different from the predicted?)
    The immunogen for ab64581 is predicted to recognise both the 70 kDa and 74 kDa isoforms of Synapsin I. Synapsin I is also known to contain a number of potential phosphorylation and glycosylation sites (SwissProt data).
  • ab64581 staining Synapsin I in rat brain (cerebellum) tissue sections by Immunohistochemistry (PFA perfusion fixed frozen sections). Tissue samples were fixed by perfusion with paraformaldehyde, permeablized with 0.01% Triton X-100, blocked with 10% serum for 2 hours at 25°C; antigen retrieval was by heat mediation in a citrate buffer. The sample was incubated with the primary antibody (1/300) at 25°C for 12 hours. An undiluted Alexa Fluor® 555-conjugated goat anti-rabbit polyclonal was used as the secondary antibody.

    See Abreview

  • Synapsin I - Synaptic Marker was immunoprecipitated using 0.5mg Mouse Brain whole tissue lysate, 5µg of Rabbit polyclonal to Synapsin I - Synaptic Marker and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, Mouse Brain whole tissue lysate lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab64581.
    Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.
    Band: 80kDa: Synapsin I - Synaptic Marker contains a number of potential glycosylation sites (SwissProt) which may explain the higher migrating band.

References for Anti-Synapsin I antibody - Synaptic Marker (ab64581)

This product has been referenced in:
  • Liu W  et al. Transplantation of olfactory ensheathing cells attenuates acute carbon monoxide poisoning-induced brain damages in rats. Neurochem Res 40:70-80 (2015). Read more (PubMed: 25370793) »
  • Rabenstein M  et al. Osteopontin mediates survival, proliferation and migration of neural stem cells through the chemokine receptor CXCR4. Stem Cell Res Ther 6:99 (2015). IHC ; Rat . Read more (PubMed: 25998490) »

See all 5 Publications for this product

Product Wall

Application Western blot
Sample Rat Tissue lysate - whole (Whole Brain)
Gel Running Conditions Reduced Denaturing (7.5)
Loading amount 15 µg
Specification Whole Brain
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jun 01 2015

I am very pleased to hear you would like to test ab5076, ab8049, ab18258, ab58802, ab64581, ab82259 and send us your results. The testing discount doesn't apply to ab4674 since this has already been tested in Rhesus monkey and IHC so we would guarantee...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample Rat Tissue sections (Brain-Cerebellum)
Specification Brain-Cerebellum
Fixative Paraformaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrate buffer
Permeabilization Yes - 0.01% Triton X-100
Blocking step Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jul 28 2012

Thank you for contacting us.
I am sorry to hear you are experiencing difficulties with our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.
I am attaching our que...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (Brain coronal sections)
Specification Brain coronal sections
Fixative Paraformaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: tris-sodium citrate buffer
Permeabilization Yes - 0.2 % triton
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Feb 24 2012

That's great to hear! I'd really appreciate it if you would take the time to submit an Abreview - this should be helpful for other customers.

Have a wonderful day, and please don't hesitate to let me know if there is anything else I can help w...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Rat Cell (Hippocampal neuron culture)
Specification Hippocampal neuron culture
Fixative Methanol
Permeabilization Yes - 0.3% TritonX in 0.1% PBS
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 24°C
Username

Dr. Ruma Raha-Chowdhury

Verified customer

Submitted Feb 09 2012

Regarding ab8, I'm very sorry to hear that it isn't working for you! This should be working in IHC on mouse tissues, so I'm not sure why it is giving you trouble. I took a look through our catalog, and it looks like ab64581 would be a good replacement....

Read More
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Monkey Tissue sections (Brain)
Specification Brain
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization No
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Username

Mr. Carl Hobbs

Verified customer

Submitted Aug 18 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Rat Tissue sections (Cerebellum)
Specification Cerebellum
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization No
Blocking step BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C
Username

Mr. Carl Hobbs

Verified customer

Submitted Aug 18 2011

1-10 of 16 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"