Recombinant Anti-Synaptophysin antibody [YE269] (ab32127)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [YE269] to Synaptophysin
- Suitable for: WB, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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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, IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat, Human
Predicted to work with: Donkey, Cow -
Immunogen
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Positive control
- WB: PC-12 and HEK-293T cell lysates; Human fetal brain, mouse brain and rat brain lysates; Neurons from iPS cells lysate. ICC/IF: PC-12 cells; Human iPS cell derived neurons, primary mouse neurons/glia, DIV14 cells. IHC-P: Human pancreas, mouse cerebral cortex, rat cerebral cortex, medullablastoma, lung neuroendocrine tumor tissues; Sheep gut tissue. IHC-Fr: Rat brain tissue.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
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.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
YE269 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- Anti-Synaptophysin antibody [YE269] - BSA and Azide free (ab187259)
- HRP Anti-Synaptophysin antibody [YE269] (ab195520)
- Alexa Fluor® 647 Anti-Synaptophysin antibody [YE269] (ab196166)
- Alexa Fluor® 488 Anti-Synaptophysin antibody [YE269] (ab196379)
- Alexa Fluor® 594 Anti-Synaptophysin antibody [YE269] (ab206868)
- Alexa Fluor® 555 Anti-Synaptophysin antibody [YE269] (ab206870)
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Isotype control
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KO cell lines
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KO cell lysates
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KO cell pellets
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
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 |
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WB | (7) |
1/20000 - 1/100000. Detects a band of approximately 34 kDa (predicted molecular weight: 34 kDa).Can be blocked with Synaptophysin peptide (ab189853).
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IHC-P | (13) |
1/400 - 1/800. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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ICC/IF | (3) |
Use a concentration of 0.1 µg/ml.
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Notes |
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WB
1/20000 - 1/100000. Detects a band of approximately 34 kDa (predicted molecular weight: 34 kDa).Can be blocked with Synaptophysin peptide (ab189853). |
IHC-P
1/400 - 1/800. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
ICC/IF
Use a concentration of 0.1 µg/ml. |
Target
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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
modificationsUbiquitinated; 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
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Database links
- Entrez Gene: 280937 Cow
- Entrez Gene: 6855 Human
- Entrez Gene: 20977 Mouse
- Entrez Gene: 24804 Rat
- Omim: 313475 Human
- SwissProt: P20488 Cow
- SwissProt: P08247 Human
- SwissProt: Q62277 Mouse
see all -
Alternative names
- Major synaptic vesicle protein p38 antibody
- MRX96 antibody
- MRXSYP antibody
see all
Images
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Immunocytochemistry/ Immunofluorescence analysis of mouse primary neuron cells labeling Synaptophysin with purified ab32127 at 1/100 (2.7µg/mL). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
Confocal scanning Z step was set as 0.3 µm followed by image processing with maximum Z projection. -
ab32127 staining Synaptophysin in primary mouse neurons/glia, DIV14 (prepared from E18 mouse hippocampal brain area, obtained from Transnetyx Tissue by BrainBits, LLC, cat.no. C57EHP) cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab32127 at 0.1µg/ml and ab192757, Mouse mono Anti-PSD95 antibody [K28/43] - Synaptic Marker. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
Also suitable in cells fixed with 100% methanol (5 min).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
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All lanes : Anti-Synaptophysin antibody [YE269] (ab32127) at 1/1000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : SYP knockout HEK-293T cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 34 kDa
Observed band size: 38 kDa why is the actual band size different from the predicted?Lanes 1- 2: Merged signal (red and green). Green - ab32127 observed at 38 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab32127 was shown to react with Syp in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab255356 (knockout cell lysate ab263862) was used. Wild-type HEK-293T and SYP knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab32127 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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ab32127 staining Synaptophysin in primary hippocampal rat neurons/glia, (obtained from Neuromics, cat. no. PC35101), DIV14. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Tween for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab32127 at 0.1µg/ml and ab192757, Mouse mono Anti-PSD95 antibody [K28/43] - Synaptic Marker. Cells were then incubated with ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488), pre-adsorbed at 1/1000 dilution (shown in green) and ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor® 594), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
Also suitable in cells fixed with 4% paraformaldehyde (10 min).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Synaptophysin antibody [YE269] (ab32127)
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.
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All lanes : Anti-Synaptophysin antibody [YE269] (ab32127) at 1/1000 dilution
Lane 1 : Wild-type HEK-293T cell lysate at 20 µg
Lane 2 : SYP knockout HEK-293T cell lysate at 20 µg
Lane 3 : Human brain tissue lysate
Performed under reducing conditions.
Predicted band size: 34 kDa
Observed band size: 38 kDa why is the actual band size different from the predicted?Lanes 1-3: Merged signal (red and green). Green - ab32127 observed at 38 kDa. Red - loading control, ab7291 observed at 50 kDa.
ab32127 Anti-Synaptophysin antibody [YE269] was shown to specifically react with Synaptophysin in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab267272 (knockout cell lysate ab257060) was used. Wild-type and Synaptophysin knockout samples were subjected to SDS-PAGE. ab32127 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti- Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti- Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.
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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.
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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
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Synaptophysin antibody [YE269] (ab32127)
Unpurified ab32127 showing positive staining in medulloblastoma tissue.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Synaptophysin antibody [YE269] (ab32127)
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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Synaptophysin antibody [YE269] (ab32127)
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.
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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. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Synaptophysin antibody [YE269] (ab32127)
Unpurified ab32127 showing positive staining in lung neuroendocrine tumor tissue.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (184)
ab32127 has been referenced in 184 publications.
- Fan XY et al. Neonatal Sevoflurane Exposure Impairs Learning and Memory by the Hypermethylation of Hippocampal Synaptic Genes. Mol Neurobiol 58:895-904 (2021). PubMed: 33052583
- Huang L et al. L-3-n-Butylphthalide improves synaptic and dendritic spine plasticity and ameliorates neurite pathology in Alzheimer's disease mouse model and cultured hippocampal neurons. Mol Neurobiol 58:1260-1274 (2021). PubMed: 33146400
- Dorigatti AO et al. Primary neuron and astrocyte cultures from postnatal Callithrix jacchus: a non-human primate in vitro model for research in neuroscience, nervous system aging, and neurological diseases of aging. Geroscience 43:115-124 (2021). PubMed: 33063253
- Kurioka T et al. Decreasing auditory input induces neurogenesis impairment in the hippocampus. Sci Rep 11:423 (2021). PubMed: 33432038
- Choi GE et al. BNIP3L/NIX-mediated mitophagy protects against glucocorticoid-induced synapse defects. Nat Commun 12:487 (2021). PubMed: 33473105