重组Anti-Syntenin抗体[EPR8102] (ab133267)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR8102] to Syntenin
- Suitable for: Flow Cyt (Intra), WB, IP, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
概述
-
产品名称
Anti-Syntenin抗体[EPR8102]
参阅全部 Syntenin 一抗 -
描述
兔单克隆抗体[EPR8102] to Syntenin -
宿主
Rabbit -
经测试应用
适用于: Flow Cyt (Intra), WB, IP, IHC-P, ICC/IFmore details -
种属反应性
与反应: Human -
免疫原
Synthetic peptide within Human Syntenin aa 1-100. The exact sequence is proprietary.
Database link: O00560 -
阳性对照
- Human fetal brain lysate, Human fetal heart lysate, Human placenta lysate, HeLa, 293T and A549 (Human lung carcinoma cell line) cell lysates; Human brain tissue
-
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.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
性能
-
形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
解离常数(KD)
KD = 1.24 x 10 -10 M Learn more about KD -
存储溶液
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
-
纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR8102 -
同种型
IgG -
研究领域
相关产品
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
-
Positive Controls
-
Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab133267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 |
---|---|---|
Flow Cyt (Intra) |
1/50.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. For unpurified use at 1/1000 - 1/10000. |
|
WB | (1) |
1/1000 - 1/10000. Detects a band of approximately 32 kDa (predicted molecular weight: 32 kDa).
|
IP |
1/10 - 1/100.
|
|
IHC-P |
1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
|
|
ICC/IF |
1/50 - 1/200.
|
说明 |
---|
Flow Cyt (Intra)
1/50. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. For unpurified use at 1/1000 - 1/10000. |
WB
1/1000 - 1/10000. Detects a band of approximately 32 kDa (predicted molecular weight: 32 kDa). |
IP
1/10 - 1/100. |
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
ICC/IF
1/50 - 1/200. |
靶标
-
功能
Seems to function as an adapter protein. In adherens junctions may function to couple syndecans to cytoskeletal proteins or signaling components. Seems to couple transcription factor SOX4 to the IL-5 receptor (IL5RA). May also play a role in vesicular trafficking. Seems to be required for the targeting of TGFA to the cell surface in the early secretory pathway. -
组织特异性
Widely expressed. Expressed in fetal kidney, liver, lung and brain. In adult highest expression in heart and placenta. -
序列相似性
Contains 2 PDZ (DHR) domains. -
翻译后修饰
Phosphorylated on tyrosine residues. -
细胞定位
Cell junction > focal adhesion. Cell junction > adherens junction. Cell membrane. Endoplasmic reticulum membrane. Nucleus. Melanosome. Cytoplasm > cytosol. Cytoplasm > cytoskeleton. Mainly membrane-associated. Localized to adherens junctions, focal adhesions and endoplasmic reticulum. Colocalized with actin stress fibers. Also found in the nucleus. Identified by mass spectrometry in melanosome fractions from stage I to stage IV. - Information by UniProt
-
数据库链接
- Entrez Gene: 6386 Human
- Omim: 602217 Human
- SwissProt: O00560 Human
- Unigene: 200804 Human
-
别名
- MDA-9 antibody
- MDA9 antibody
- Melanoma differentiation-associated protein 9 antibody
see all
图片
-
Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: Syntenin knockout HAP1 whole cell lysate (20 µg)
Lane 3: A549 whole cell lysate (20 µg)
Lane 4: HeLa whole cell lysate (20 µg)Lanes 1 - 4: Merged signal (red and green). Green - ab133267 observed at 32 kDa. Red - loading control, ab130007, observed at 130 kDa.
ab133267 was shown to recognize Syntenin in wild-type HAP1 cells as signal was lost at the expected MW in Syntenin knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and Syntenin knockout samples were subjected to SDS-PAGE. ab133267 and ab130007 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Syntenin antibody [EPR8102] (ab133267)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human hepatocellular carcinoma tissue labeling Syntenin with purified ab133267 at 1/50 dilution. Heat mediated antigen retrieval was performed using EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin
-
Anti-Syntenin antibody [EPR8102] (ab133267) at 1/2000 dilution (purified) + A549 (Human lung carcinoma cell line) whole cell lysate at 15 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 32 kDa
Observed band size: 32 kDaBlocking and dilution buffer: 5% NFDM/TBST
-
Intracellular Flow Cytometry analysis ofHeLa (Human epithelial cell line from cervix adenocarcinoma)labeling Syntenin with purifiedab133267 at 1/50 (red).Cells were fixed with 4% paraformaldehyde. A goat anti rabbit IgG (Alexa Fluor® 488) 1/2000 was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.
-
ab133267 (purified) at 1/40 immunoprecipitating Syntenin in HeLa whole cell lysate.
Lane 1 (input): HeLa whole cell lysate (10µg)
Lane 2 (+): ab133267 + HeLa whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab133267 in HeLa whole cell lysate.
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Syntenin antibody [EPR8102] (ab133267)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human cerebral cortex tissue labeling Syntenin with purified ab133267 at 1/50 dilution. Heat mediated antigen retrieval was performed using EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
-
All lanes : Anti-Syntenin antibody [EPR8102] (ab133267) at 1/2000 dilution (purified)
Lane 1 : Human placenta lysate
Lane 2 : Human fetal brain lysate
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution
Predicted band size: 32 kDa
Observed band size: 32 kDaBlocking and dilution buffer: 5% NFDM/TBST
-
Anti-Syntenin antibody [EPR8102] (ab133267) at 1/10000 dilution (purified) + HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Secondary
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution
Predicted band size: 32 kDa
Observed band size: 32 kDaBlocking and dilution buffer: 5% NFDM/TBST
-
ab133267 staining Syntenin in HeLa (human cervix adenocarcinoma) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% Paraformaldehyde and permeabilized with 0.1% Triton X-100. Samples were incubated with primary antibody at a dilution of 1/500. A goat anti rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at a dilution of 1/1000. DAPI was used as a nuclear counterstain.
Negative control 1: PBS only.
-
Overlay histogram showing SHSY-5Y cells stained with unpurified ab133267 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab133267, 1/10000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in SHSY-5Y cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
-
All lanes : Anti-Syntenin antibody [EPR8102] (ab133267) at 1/1000 dilution (unpurified)
Lane 1 : Human fetal brain lysate
Lane 2 : 293T cell lysate
Lane 3 : Human fetal heart lysate
Lane 4 : HeLa cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat anti-rabbit HRP conjugated antibody at 1/2000 dilution
Predicted band size: 32 kDa
Observed band size: 32 kDa -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Syntenin antibody [EPR8102] (ab133267)
Immunohistochemical analysis of Syntenin in paraffin embedded Human brain tissue, using unpurified ab133267 at a 1/50 dilution.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
实验方案
数据表及文件
-
SDS download
-
Datasheet download
文献 (88)
ab133267 被引用在 88 文献中.
- Marie PP et al. Accessory ESCRT-III proteins are conserved and selective regulators of Rab11a-exosome formation. J Extracell Vesicles 12:e12311 (2023). PubMed: 36872252
- Powell BH et al. miR-210 Expression Is Strongly Hypoxia-Induced in Anaplastic Thyroid Cancer Cell Lines and Is Associated with Extracellular Vesicles and Argonaute-2. Int J Mol Sci 24:N/A (2023). PubMed: 36901936
- Palamà MEF et al. Xeno-free cultured mesenchymal stromal cells release extracellular vesicles with a "therapeutic" miRNA cargo ameliorating cartilage inflammation in vitro. Theranostics 13:1470-1489 (2023). PubMed: 37056573
- Andriolo G et al. Methodologies for Scalable Production of High-Quality Purified Small Extracellular Vesicles from Conditioned Medium. Methods Mol Biol 2668:69-98 (2023). PubMed: 37140791
- Dhondt B et al. Benchmarking blood collection tubes and processing intervals for extracellular vesicle performance metrics. J Extracell Vesicles 12:e12315 (2023). PubMed: 37202906