重组Anti-Synaptophysin抗体[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
概述
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产品名称
Anti-Synaptophysin抗体[YE269]
参阅全部 Synaptophysin 一抗 -
描述
兔单克隆抗体[YE269] to Synaptophysin -
宿主
Rabbit -
经测试应用
适用于: WB, IHC-P, ICC/IFmore details -
种属反应性
与反应: Mouse, Rat, Human
预测可用于: Donkey, Cow -
免疫原
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阳性对照
- 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.
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常规说明
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.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
存储溶液
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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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
YE269 -
同种型
IgG -
研究领域
相关产品
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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)
- Anti-Synaptophysin antibody [YE269] - Mouse IgG1 (Chimeric) (ab309493)
- Anti-Synaptophysin antibody [YE269] - Mouse IgG1 (Chimeric) - BSA and Azide free (ab309494)
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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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Recombinant Protein
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab32127于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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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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说明 |
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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. |
靶标
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功能
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. -
组织特异性
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. -
疾病相关
Mental retardation, X-linked, SYP-related -
序列相似性
Belongs to the synaptophysin/synaptobrevin family.
Contains 1 MARVEL domain. -
结构域
The calcium-binding activity is thought to be localized in the cytoplasmic tail of the protein. -
翻译后修饰
Ubiquitinated; mediated by SIAH1 or SIAH2 and leading to its subsequent proteasomal degradation. -
细胞定位
Cytoplasmic vesicle, secretory vesicle, synaptic vesicle membrane. Cell junction, synapse, synaptosome. - Information by UniProt
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数据库链接
- 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 -
别名
- Major synaptic vesicle protein p38 antibody
- MRX96 antibody
- MRXSYP antibody
see all
图片
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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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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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Unpurified ab32127 showing positive staining in medulloblastoma tissue.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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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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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. -
Unpurified ab32127 showing positive staining in lung neuroendocrine tumor tissue.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
实验方案
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.
数据表及文件
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SDS download
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Datasheet download
文献 (255)
ab32127 被引用在 255 文献中.
- Li X et al. Involvement of Paired Immunoglobulin-Like Receptor B in Cognitive Dysfunction Through Hippocampal-Dependent Synaptic Plasticity Impairments in Mice Subjected to Chronic Sleep Restriction. Mol Neurobiol 60:1132-1149 (2023). PubMed: 36417104
- Dong D et al. VX-765 Alleviates β-Amyloid Deposition and Secondary Degeneration in the Ipsilateral Hippocampus and Ameliorates Cognitive Decline after Focal Cortical Infarction in Rats. J Mol Neurosci 72:2389-2397 (2022). PubMed: 36441377
- Li ZD et al. The divergent effects of astrocyte ceruloplasmin on learning and memory function in young and old mice. Cell Death Dis 13:1006 (2022). PubMed: 36443285
- Zhang PN et al. Integrated Network Pharmacology Analysis and Experimental Validation to Investigate the Molecular Mechanism of Triptolide in the Treatment of Membranous Nephropathy. Drug Des Devel Ther 16:4061-4076 (2022). PubMed: 36448035
- Hu T et al. Astrocyte derived TSP2 contributes to synaptic alteration and visual dysfunction in retinal ischemia/reperfusion injury. Cell Biosci 12:196 (2022). PubMed: 36471420