- 数据表
- 具体的参考文献 (17)
- 实验方案
概述
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产品名称Anti-alpha Adaptin抗体[AP6]
参阅全部 alpha Adaptin 一抗 -
描述小鼠单克隆抗体[AP6] to alpha Adaptin
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宿主Mouse
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特异性Detects assembly polypeptide 2 (AP2) It recognizes the products of both alpha-adaptin genes, alpha A and alpha C as well as an alternatively spliced isoform of alpha A found in neurons.
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经测试应用适用于: ICC/IF, ICC, Inhibition Assay, Electron Microscopy, ELISA, Blocking, IP, WB, IHC-FoFr, Flow Cytmore details
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种属反应性与反应: Mouse, Rat, Hamster, Cow, Human, Drosophila melanogaster
预测可用于: Non human primates
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免疫原
Other Immunogen Type corresponding to alpha Adaptin. Purified adaptors.
性能
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形式Liquid
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存放说明Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
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存储溶液Preservative: 0.05% Sodium azide
Constituent: PBS -
Concentration information loading... -
纯度Affinity purified
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克隆单克隆
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克隆编号AP6
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同种型IgG1
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研究领域
相关产品
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Compatible Secondaries
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Conjugation kits
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Isotype control
应用
Our Abpromise guarantee covers the use of ab2730 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| 应用 | Ab评论 | 说明 |
|---|---|---|
| ICC/IF | Use a concentration of 5 µg/ml. | |
| ICC | Use at an assay dependent concentration. | |
| Inhibition Assay | Use at an assay dependent concentration. | |
| Electron Microscopy | Use at an assay dependent concentration. PubMed: 23372794 | |
| ELISA | Use at an assay dependent concentration. | |
| Blocking | Use at an assay dependent concentration. | |
| IP | Use at an assay dependent concentration. | |
| WB | 1/1000. This antibody detects an ~100 kDa doublet representing the two isoforms of alpha-adaptin in rat brain, though alpha C is the dominant form seen. Microinjection of cells with this antibody results in variable blockade of endocytosis. | |
| IHC-FoFr | Use at an assay dependent concentration. | |
| Flow Cyt | Use 1µg for 106 cells. ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
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靶标
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相关性Clathrin mediated endocytosis is the pathway by which many receptors for nutrients and hormones are internalized to be recycled or down regulated. During formation of clathrin coated membranes, clathrin co assembles with heterotetrameric molecules known as assembly polypeptides (APs) or adaptors which form a layer of protein coat between the clathrin lattice and the membrane. There are two characterized adaptors AP1 and AP2. AP1 is associated with clathrin coated vesicles at the trans Golgi network and AP2 is associated with the endocytic clathrin coated vesicles at the plasma membrane and has been shown to specifically interact with Shc and EGF receptor. AP2 is composed of four subunits, two separate 100 kDa gene products with similar domain structures (alpha and beta adaptin) and a 50 and 17 kDa subunit. There are two alpha adaptin genes, alpha A and alpha C which have a tissue specific pattern of expression.
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细胞定位Cytoplasmic
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数据库链接
- Entrez Gene: 515216 Cow
- Entrez Gene: 784636 Cow
- Entrez Gene: 160 Human
- Entrez Gene: 161 Human
- Entrez Gene: 11771 Mouse
- Entrez Gene: 11772 Mouse
- Entrez Gene: 308578 Rat
- Entrez Gene: 81637 Rat
see all -
别名
- 100 kDa coated vesicle protein A antibody
- 100 kDa coated vesicle protein C antibody
- Adapter related protein complex 2 subunit alpha 1 antibody
see all
图片
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![Immunocytochemistry/ Immunofluorescence - Anti-alpha Adaptin antibody [AP6] (ab2730)](http://www.abcam.cn/ps/products/2/ab2730/Images/ab2730-200603-Antialpha-Adaptin-antibody-AP6.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-alpha Adaptin antibody [AP6] (ab2730)Immunofluorescent analysis of alpha Adaptin using alpha Adaptin Monoclonal Antibody (AP6) (ab2730) shows staining in U251 Cells. alpha Adaptin (green) F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with an antibody recognizing alpha Adaptin (ab2730) at a dilution of 1:20 over night at 4C and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification. -
![Immunocytochemistry/ Immunofluorescence - Anti-alpha Adaptin antibody [AP6] (ab2730)](http://www.abcam.cn/ps/products/2/ab2730/Images/ab2730-200602-Antialpha-Adaptin-antibody-AP6.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-alpha Adaptin antibody [AP6] (ab2730)Immunofluorescent analysis of alpha Adaptin using alpha Adaptin Monoclonal Antibody (AP6) (ab2720) shows staining in MCF-7 Cells. alpha Adaptin (green) F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with an antibody recognizing alpha Adaptin (ab2730) at a dilution of 1:20 over night at 4 C and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification. -
![Immunocytochemistry/ Immunofluorescence - Anti-alpha Adaptin antibody [AP6] (ab2730)](http://www.abcam.cn/ps/products/2/ab2730/Images/ab2730-200601-Antialpha-Adaptin-antibody-AP6.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-alpha Adaptin antibody [AP6] (ab2730)Immunofluorescent analysis of alpha Adaptin using alpha Adaptin Monoclonal Antibody (AP6) (ab2730) shows staining in Hela Cells. alpha Adaptin (green) F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with an antibody recognizing alpha Adaptin (ab2720) at a dilution of 1:20 over night at 4 C and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification. -
![Immunocytochemistry/ Immunofluorescence - Anti-alpha Adaptin antibody [AP6] (ab2730)](http://www.abcam.cn/ps/products/2/ab2730/Images/alpha-Adaptin-Primary-antibodies-ab2730-1.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-alpha Adaptin antibody [AP6] (ab2730)ICC/IF image of ab2730 stained MCF7 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab2730, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. -
![Flow Cytometry - Anti-alpha Adaptin antibody [AP6] (ab2730)](http://www.abcam.cn/ps/products/2/ab2730/Images/alpha-Adaptin-Primary-antibodies-ab2730-3.jpg)
Flow Cytometry - Anti-alpha Adaptin antibody [AP6] (ab2730)Overlay histogram showing MCF7 cells stained with ab2730 (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 (ab2730, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was a goat anti-mouse DyLight® 488 (IgG; H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
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![Western blot - Anti-alpha Adaptin antibody [AP6] (ab2730)](http://www.abcam.cn/ps/products/2/ab2730/Images/alpha-Adaptin-Primary-antibodies-ab2730-6.jpg)
Western blot - Anti-alpha Adaptin antibody [AP6] (ab2730)Image from Li Y et al, J Biol Chem. 2012 Jan 2;287(1):523-30. Epub 2011 Nov 11, Fig 2. DOI 10.1074/jbc.M111.306183 January 2, 2012 The Journal of Biological Chemistry, 287, 523-530.ab2730 used in Western Blot at a 1/1000 dilution.
Sucrose density gradient fractionation of detergent resistant cell lysates from WT and DHHC5-GT murine neuronal stem cells. The isolation of detergent-resistant membranes was performed according to a previously published protocol. NSCs growing in growth medium (~300 µl of packed cells) were harvested and washed three times with ice-cold PBS. The cells were gently resuspended by pipetting up and down in 700 µl of 1% Triton X-100 in HEPES buffer (25 mm HEPES-HCl, pH 6.5, 150 mm NaCl, 1 mm EDTA, and protease inhibitor mixture), homogenized using a Teflon-coated Dounce homogenizer (30 strokes), then incubated on ice for 30 min, and fractionated on a sucrose density gradient. The fractions were collected from top (fraction 1) to bottom (fraction 13) and analyzed by immunoblotting. The results shown were from one of two independent experiments giving similar results. a-Adaptin is a marker for clathrin-coated vesicles.
实验方案
文献
This product has been referenced in:
- Evergren E et al. Eps15R and clathrin regulate EphB2-mediated cell repulsion. Traffic 19:44-57 (2018). Read more (PubMed: 28972287) »
- Humphries AC et al. Cdc42 and the Rho GEF intersectin-1 collaborate with Nck to promote N-WASP-dependent actin polymerisation. J Cell Sci 127:673-85 (2014). Read more (PubMed: 24284073) »
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