Recombinant
RabMAb

Anti-Synaptophysin antibody [YE269] (ab32127)

Overview

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

    corresponding to Human Synaptophysin aa 250-350 (C terminal).
    (Peptide available as ab189853)

  • Positive control
    • WB: PC12 cell lysate, human fetal brain, mouse brain, rat brain IF: PC12 cells. IHC: Human pancreas, mouse cerebral cortex, rat cerebral cortex, medullablastoma, lung neuroendocrine tumor
  • General notes

     

     

    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 protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

IHC-FoFr Use at an assay dependent concentration. PubMed: 25040426

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

  • Immunofluorescent staining of PC-12 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 counter stained 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.

  • Unpurified ab32127 showing positive staining in Medulloblastoma tissue.

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

    Lane 1 : Mouse brain
    Lane 2 : Rat brain

    Secondary
    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 buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • 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 counter-stained wirh 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 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 counter-stained wirh 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 counter-stained wirh 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) + PC12 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:
  • Jiang T  et al. TREM2 Overexpression has No Improvement on Neuropathology and Cognitive Impairment in Aging APPswe/PS1dE9 Mice. Mol Neurobiol 54:855-865 (2017). WB ; Mouse . Read more (PubMed: 26780455) »
  • Bellissimo T  et al. Thymic Epithelial Tumors phenotype relies on miR-145-5p epigenetic regulation. Mol Cancer 16:88 (2017). WB ; Human . Read more (PubMed: 28486946) »

See all 35 Publications for this product

Customer reviews and Q&As

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
Username

Abcam user community

Verified customer

Submitted Sep 27 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry free floating
Sample
Mouse Tissue sections (Brain)
Specification
Brain
Username

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
Username

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
Username

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
Username

Abcam user community

Verified customer

Submitted Jun 05 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Horse Cell (Cerebellum)
Permeabilization
No
Specification
Cerebellum
Fixative
None
Username

Stephen Cahalan

Verified customer

Submitted May 12 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Horse Tissue sections (Adrenal gland)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate buffer
Permeabilization
No
Specification
Adrenal gland
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 20% · Temperature: 22°C
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Feb 06 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Dog Tissue sections (pancreas)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: CC1
Permeabilization
No
Specification
pancreas
Fixative
formalin 4%
Username

Dr. Emmanuel Bouchaert

Verified customer

Submitted Jan 17 2017

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)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: TE pH9
Permeabilization
No
Specification
Brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Fixative
10% Normal buffered formalin
Username

Abcam user community

Verified customer

Submitted Dec 05 2016

Application
IHC - Wholemount
Sample
Mouse Tissue (Diaphragm)
Specification
Diaphragm
Username

Abcam user community

Verified customer

Submitted Oct 25 2016

1-10 of 28 Abreviews or Q&A

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