Overview

  • Product nameAnti-alpha Adaptin antibody [AP6]
    See all alpha Adaptin primary antibodies
  • Description
    Mouse monoclonal [AP6] to alpha Adaptin
  • SpecificityDetects 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.
  • Tested applicationsSuitable for: ICC/IF, ICC, Inhibition Assay, Electron Microscopy, ELISA, Blocking, IP, WB, IHC-FoFr, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Hamster, Cow, Human, Drosophila melanogaster
    Predicted to work with: Non Human Primates
  • Immunogen

    Other Immunogen Type corresponding to alpha Adaptin. Purified adaptors.

Properties

  • FormLiquid
  • Storage instructionsShipped 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.
  • Storage bufferPreservative: 0.05% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • PurityIgG fraction
  • ClonalityMonoclonal
  • Clone numberAP6
  • IsotypeIgG1
  • Research areas

Applications

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.

Application Abreviews Notes
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.

Target

  • RelevanceClathrin 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.
  • Cellular localizationCytoplasmic
  • Database links
  • Alternative names
    • 100 kDa coated vesicle protein A antibody
    • 100 kDa coated vesicle protein C antibody
    • Adapter related protein complex 2 subunit alpha 1 antibody
    • Adapter related protein complex 2 subunit alpha 2 antibody
    • Adaptin alpha A antibody
    • Adaptin alpha B antibody
    • Adaptin, alpha B antibody
    • Adaptor protein complex AP 2 alpha 1 subunit antibody
    • Adaptor protein complex AP 2 alpha 2 subunit antibody
    • Adaptor protein complex AP 2 subunit alpha 1 antibody
    • Adaptor related protein complex 2 alpha 1 subunit antibody
    • Adaptor related protein complex 2 alpha 2 subunit antibody
    • ADTAA antibody
    • ADTAB antibody
    • Alpha adaptin A antibody
    • Alpha adaptin C antibody
    • Alpha adaptin C; Huntingtin interacting protein J antibody
    • Alpha1 adaptin antibody
    • Alpha2 adaptin antibody
    • AP 2 complex subunit alpha 1 antibody
    • AP 2 complex subunit alpha 2 antibody
    • AP2 alpha antibody
    • AP2A1 antibody
    • AP2A2 antibody
    • CLAPA1 antibody
    • CLAPA2 antibody
    • Clathrin assembly protein complex 2 alpha A large chain antibody
    • Clathrin assembly protein complex 2 alpha C large chain antibody
    • Clathrin associated/assembly/adaptor protein large alpha 1 antibody
    • Clathrin associated/assembly/adaptor protein large alpha 2 antibody
    • HIP 9 antibody
    • HIP9 antibody
    • Huntingtin interacting protein antibody
    • Huntingtin interacting protein HYPJ antibody
    • Huntingtin interacting protein J antibody
    • Huntingtin yeast partner J antibody
    • HYPJ antibody
    • Plasma membrane adaptor HA2/AP2 adaptin alpha A subunit antibody
    • Plasma membrane adaptor HA2/AP2 adaptin alpha C subunit antibody
    see all

Anti-alpha Adaptin antibody [AP6] images

  • 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.

  • 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.

References for Anti-alpha Adaptin antibody [AP6] (ab2730)

This product has been referenced in:
  • 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) »
  • Grassart A  et al. Actin and dynamin2 dynamics and interplay during clathrin-mediated endocytosis. J Cell Biol 205:721-35 (2014). Read more (PubMed: 24891602) »

See all 15 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (undifferentiated Human ES cells on MEFs)
Permeabilization Yes - in blocking buffer
Specification undifferentiated Human ES cells on MEFs
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative Paraformaldehyde
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Submitted Feb 01 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (Undifferentiated Human ES cells on MEFs)
Gel Running Conditions Non-reduced Denaturing (12.5%)
Loading amount 20 µg
Specification Undifferentiated Human ES cells on MEFs
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: RT°C
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Submitted Jan 25 2016

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HEPG2)
Specification HEPG2
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: rt°C
Fixative Paraformaldehyde
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Submitted Nov 18 2015

Application Western blot
Sample Human Cell lysate - whole cell (HEPG2)
Gel Running Conditions Reduced Denaturing (12.5)
Loading amount 20 µg
Specification HEPG2
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 16°C
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Submitted Nov 13 2015

Application Immunocytochemistry/ Immunofluorescence
Blocking step 0.1% BSA and 1% serum as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 16°C
Sample Mouse Cell (mouse embryonic stem cell)
Specification mouse embryonic stem cell
Permeabilization Yes - 0.1% triton x
Fixative methanol/acetone
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Submitted May 19 2014

Application Western blot
Loading amount 100000 cells
Gel Running Conditions Reduced Denaturing (10% gel)
Sample Mouse Cell lysate - whole cell (mouse embryonic stem cells)
Specification mouse embryonic stem cells
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 16°C
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Submitted Jan 09 2014

Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Blocking step Serum as blocking agent for 20 minute(s) · Concentration: 10% · Temperature: 22°C
Sample Human Cell (HeLa)
Specification HeLa
Permeabilization Yes - 0.2% Triton in PBS
Fixative Formaldehyde
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Submitted Mar 05 2013

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Application Western blot
Sample Mouse Cell lysate - whole cell (mouse neuro2A)
Loading amount 90 µg
Specification mouse neuro2A
Gel Running Conditions Non-reduced Non-Denaturing (Native) (8% running gel)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted Dec 21 2012

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