Overview

  • Product name
    Anti-Synaptophysin antibody [YE269]
    See all Synaptophysin primary antibodies
  • Description
    Rabbit monoclonal [YE269] to Synaptophysin
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, ICC/IF, IHC-Fr, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Donkey, Human
    Predicted to work with: Cow
  • Immunogen

    Synthetic peptide within Human Synaptophysin aa 250-350 (C terminal). The exact sequence is proprietary.
    (Peptide available as ab189853)

  • Positive control
    • WB: PC-12 cell lysate; Human fetal brain, mouse brain and rat brain lysates; Neurons from iPS cells lysate. ICC/IF: PC-12 cells; Human iPS cell derived neurons. IHC-P: Human pancreas, mouse cerebral cortex, rat cerebral cortex, medullablastoma, lung neuroendocrine tumor tissues. IHC-Fr: Rat brain tissue.
  • General notes

     

     

    Abcam recommended secondaries - Goat Anti-Rabbit HRP (ab205718) and Goat Anti-Rabbit Alexa Fluor® 488 (ab150077).

    See other anti-rabbit secondary antibodies that can be used with this antibody.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab32127 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 1/20000 - 1/100000. Detects a band of approximately 34 kDa (predicted molecular weight: 34 kDa).Can be blocked with Synaptophysin peptide (ab189853).
ICC/IF 1/50 - 1/250.

 

IHC-Fr Use at an assay dependent concentration.
IHC-P 1/400 - 1/800. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

Target

  • Function
    Possibly involved in structural functions as organizing other membrane components or in targeting the vesicles to the plasma membrane. Involved in the regulation of short-term and long-term synaptic plasticity.
  • Tissue specificity
    Characteristic of a type of small (30-80 nm) neurosecretory vesicles, including presynaptic vesicles, but also vesicles of various neuroendocrine cells of both neuronal and epithelial phenotype.
  • Involvement in disease
    Mental retardation, X-linked, SYP-related
  • Sequence similarities
    Belongs to the synaptophysin/synaptobrevin family.
    Contains 1 MARVEL domain.
  • Domain
    The calcium-binding activity is thought to be localized in the cytoplasmic tail of the protein.
  • Post-translational
    modifications
    Ubiquitinated; mediated by SIAH1 or SIAH2 and leading to its subsequent proteasomal degradation.
  • Cellular localization
    Cytoplasmic vesicle, secretory vesicle, synaptic vesicle membrane. Cell junction, synapse, synaptosome.
  • Information by UniProt
  • Database links
  • Alternative names
    • Major synaptic vesicle protein p38 antibody
    • MRX96 antibody
    • MRXSYP antibody
    • Syn p38 antibody
    • Synaptophysin antibody
    • Syp antibody
    • SYPH antibody
    • SYPH_HUMAN antibody
    • SypI antibody
    see all

Images

  • All lanes : Anti-Synaptophysin antibody [YE269] (ab32127) at 1/100000 dilution (purified)

    Lane 1 : Mouse brain
    Lane 2 : Rat brain

    Secondary
    All lanes : HRP goat anti-rabbit at 1/1000 dilution

    Predicted band size: 34 kDa
    Observed band size: 38 kDa
    why is the actual band size different from the predicted?



    Blocking/Dilution buffer: 5% NFDM/TBST

  • Immunohistochemical staining of paraffin embedded mouse cerebral cortex with purified ab32127 at a dilution of 1/400.

    A pre-diluted HRP polymer for rabbit/mouse IgG was used as the secondary antibody and the sample was counterstained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0.

    PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

  • ab32127 staining synaptophysin in human iPS cell derived neurons by immunocytochemistry/immmunofluorescence.

    Samples were fixed with paraformaldehyde and blocked with 1% serum for 30 minutes at room temperature. Samples were incubated with primary antibody at 1/250 dilution for 1 hour. ab150061 was used as the secondary antibody at 1/250 dilution. 

    See Abreview

  • Immunofluorescent staining of PC-12 (rat adrenal gland pheochromocytoma cell line) cells (fixed in 4% PFA, permeabilized with 0.1% Triton X 100) using purified ab32127 at a dilution of 1/50.

    An Alexa Fluor® 488 goat anti-rabbit antibody was used as the secondary at a dilution of 1/500 and the cells were counterstained with DAPI.

    The negative control is shown in the bottom right hand panel - for the negative control, Alex Fluor® 594 goat anti-mouse was used at a dilution of 1/500.

  • ab32127 staining Synaptophysin in rat brain tissue sections by Immunohistochemistry (IHC-Fr - frozen sections).

    Tissue was fixed with formaldehyde, permeabilized with 0.2% Triton X-100 and blocked with 5% BSA for 2 hours at 25°C. Samples were incubated with primary antibody at 1/300 dilution for 10 hours at 4°C. An Alexa Fluor® 555 rabbit polyclonal was used as a secondary antibody at 1/400 dilution.

    See Abreview

  • Unpurified ab32127 showing positive staining in medulloblastoma tissue.

  • Anti-Synaptophysin antibody [YE269] (ab32127) at 1/20000 dilution + Neurons from iPS cells at 20 µg

    Secondary
    goat anti rabbit IgG HRP at 1/2000 dilution

    Predicted band size: 34 kDa



    Samples were incubated with primary antibody for 16 hours at 4°C in 5% milk in TBST.

    Blocking buffer: 5% milk for 30 minutes at room temperature. 

     

    See Abreview

  • Immunohistochemical staining of paraffin embedded rat cerebral cortex with purified ab32127 at a dilution of 1/400.

    A pre-diluted HRP polymer for rabbit/mouse IgG was used as the secondary antibody and the sample was counterstained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0.

    PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

  • Immunohistochemical staining of paraffin embedded human pancreas with purified ab32127 at a dilution of 1/400.

    A pre-diluted HRP polymer for rabbit/mouse IgG was used as the secondary antibody and the sample was counterstained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0.

    PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

  • Anti-Synaptophysin antibody [YE269] (ab32127) at 1/100000 dilution (purified) + Human fetal brain at 20 µg

    Secondary
    HRP goat anti-rabbit (H+L) at 1/1000 dilution

    Predicted band size: 34 kDa
    Observed band size: 38 kDa why is the actual band size different from the predicted?

  • Anti-Synaptophysin antibody [YE269] (ab32127) at 1/10000 dilution (unpurified) + PC-12 (rat adrenal gland pheochromocytoma cell line) cell lysate

    Predicted band size: 34 kDa
    Observed band size: 34 kDa
    Additional bands at: 21 kDa, 22 kDa. We are unsure as to the identity of these extra bands.

  • Unpurified ab32127 showing positive staining in lung neuroendocrine tumor tissue.

References

This product has been referenced in:
  • Velazquez R  et al. Maternal choline supplementation ameliorates Alzheimer's disease pathology by reducing brain homocysteine levels across multiple generations. Mol Psychiatry N/A:N/A (2019). Read more (PubMed: 30622336) »
  • Guo H  et al. ONECUT2 is a driver of neuroendocrine prostate cancer. Nat Commun 10:278 (2019). Read more (PubMed: 30655535) »
See all 64 Publications for this product

Customer reviews and Q&As

Filter by Application

Filter by Species

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1-10 of 29 Abreviews

Application
Western blot
Sample
Rat Cell lysate - whole cell (primary culture hippocampus cell)
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (15% SDS-PAGE)
Loading amount
15 µg
Treatment
RIPA & 1%Protease inhibitor for 15min on ice
Specification
primary culture hippocampus cell
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Nov 06 2018

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Cynomolgus monkey Tissue sections (Brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate
Permeabilization
No
Specification
Brain
Blocking step
Goat Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Oct 31 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Sheep Tissue sections (Gut (ileum))
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: CitricAcid buffer
Permeabilization
No
Specification
Gut (ileum)
Blocking step
NGS/PBS as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative
Formaldehyde

Cathelijne Heymans

Verified customer

Submitted Oct 11 2018

Abcam has not validated the combination of species/application used in this Abreview.
Application
IHC - Wholemount
Sample
Xenopus laevis Tissue (hindlimb muscles)
Specification
hindlimb muscles

Abcam user community

Verified customer

Submitted Oct 09 2018

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (IPS-derived Neurons)
Specification
IPS-derived Neurons
Blocking step
5% GS as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative
Methanol

Abcam user community

Verified customer

Submitted Sep 27 2017

Application
Immunohistochemistry free floating
Sample
Mouse Tissue sections (Brain)
Specification
Brain

Abcam user community

Verified customer

Submitted Aug 30 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Rat Tissue sections (Brain Tissue)
Permeabilization
Yes - 0.2% Triton X-100
Specification
Brain Tissue
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Jul 03 2017

Application
Western blot
Sample
Mouse Tissue lysate - whole (Whole brain tissue lysate)
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (3-8% Tris-Acetate)
Loading amount
20 µg
Specification
Whole brain tissue lysate
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Jun 05 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Whole brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate
Permeabilization
Yes - 0.05% Tween-20
Specification
Whole brain
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Jun 05 2017

Application
Immunocytochemistry/ Immunofluorescence
Sample
Horse Cell (Cerebellum)
Permeabilization
No
Specification
Cerebellum
Fixative
None

Stephen Cahalan

Verified customer

Submitted May 12 2017

1-10 of 29 Abreviews

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