Key features and details
- Mouse monoclonal [AP6] to alpha Adaptin
- Suitable for: ICC/IF, WB, Flow Cyt
- Reacts with: Mouse, Rat, Hamster, Cow, Human
- Isotype: IgG1
製品名Anti-alpha Adaptin antibody [AP6]
alpha Adaptin 一次抗体 製品一覧
製品の詳細Mouse monoclonal [AP6] to alpha Adaptin
特異性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.
アプリケーション適用あり: ICC/IF, WB, Flow Cytmore details
種交差性交差種: Mouse, Rat, Hamster, Cow, Human
交差が予測される動物種: Non human primates
Other Immunogen Type corresponding to alpha Adaptin. Purified adaptors.
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In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
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保存方法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.
バッファーPreservative: 0.05% Sodium azide
Concentration information loading...
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.
|ICC/IF||Use a concentration of 5 µg/ml.|
|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.|
|Flow Cyt||Use 1µg for 106 cells.
ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
関連性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.
- 100 kDa coated vesicle protein A antibody
- 100 kDa coated vesicle protein C antibody
- Adapter related protein complex 2 subunit alpha 1 antibody
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.
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.
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.
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.
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.
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.
ab2730 は 19 報の論文で使用されています。
- Wagner W et al. Myosin VI Drives Clathrin-Mediated AMPA Receptor Endocytosis to Facilitate Cerebellar Long-Term Depression. Cell Rep 28:11-20.e9 (2019). PubMed: 31269433
- Evergren E et al. Eps15R and clathrin regulate EphB2-mediated cell repulsion. Traffic 19:44-57 (2018). 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). PubMed: 24284073
- Grassart A et al. Actin and dynamin2 dynamics and interplay during clathrin-mediated endocytosis. J Cell Biol 205:721-35 (2014). PubMed: 24891602
- Bitsikas V et al. Clathrin-independent pathways do not contribute significantly to endocytic flux. Elife 3:e03970 (2014). PubMed: 25232658