Overview

  • Product nameAnti-Clathrin antibody [X22]
    See all Clathrin primary antibodies
  • Description
    Mouse monoclonal [X22] to Clathrin
  • SpecificityDetects clathrin heavy chain.
  • Tested applicationsSuitable for: ICC/IF, ICC, ELISA, Blocking, Flow Cyt, Immunomicroscopy, IP, WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Cow, Dog, Human, Pig, Xenopus laevis, Bird, African Green Monkey
  • Immunogen

    Full length native protein (purified) corresponding to Human Clathrin. Purified human brain clathrin heavy chain.

  • EpitopeElectron microscopy and proteolysis mapping demonstrate that binding occurs towards the central hub of the triskelion, N-terminal to the light chain binding regions.
  • Positive control
    • bovine brain extract

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 numberX22
  • IsotypeIgG1
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab2731 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 1/1000.
ICC Use at an assay dependent concentration.
ELISA Use at an assay dependent concentration.
Blocking Use at an assay dependent concentration.
Flow Cyt 1/200.
Immunomicroscopy Use at an assay dependent concentration.
IP Use at an assay dependent concentration. Immunoprecipitation of triskelions with this antibody allows both the clathrin heavy chain and the associated light chain polypeptides to be examined.
WB 1/500. Detects a band of approximately 180 kDa.
IHC-P 1/100.

Target

  • RelevanceMembrane vesicle formation is a process required for the endocytosis and biosynthesis of various secreted and membrane bound proteins. Clathrin is a protein which assembles into a polyhedral network on the cell membrane as the membrane invaginates, forming a coated pit which is essential to endocytosis. Clathrin is composed of three polypeptides, a 180 kDa heavy chain and two 32-38 kDa light chains which combine to create a distinct three-legged triskelion. It is this morphology which allows Clathrin to form its unique polyhedral network.
  • Cellular localizationCytoplasmic face of coated pits and vesicles.
  • Database links
  • Alternative names
    • Adapter related protein complex 1 beta 1 subunit antibody
    • Adaptor Protein Complex AP1 Beta 1 Subunit antibody
    • ADTB1 antibody
    • AP 1 complex subunit beta 1 antibody
    • AP105A antibody
    • AP1B1 antibody
    • BAM22 antibody
    • Beta Adaptin 1 antibody
    • Beta Prime Adaptin antibody
    • CLAPB2 antibody
    • Clathrin Assembly Protein Complex 1 Beta Large Chain antibody
    • Clathrin heavy chain 1 antibody
    • CLH 17 antibody
    • CLH17 antibody
    • CLTC antibody
    • CLTCL2 antibody
    • Golgi Adaptor HA1/AP1 Adaptin Beta Subunit antibody
    • KIAA0034 antibody
    • Plasma Membrane Adaptor HA2/AP2 Adaptor Beta Subunit antibody
    see all

Anti-Clathrin antibody [X22] images

  • Immunocytochemistry/Immunofluorescence analysis of Clathrin shows staining in HeLa cells. Clathrin, Heavy chain staining (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 ab2731 (1:200) over night at 4°C, washed with PBS and incubated with a DyLight-488 conjugated goat anti-mouse secondary antibody. Images were taken at 60X magnification.

  • Immunocytochemistry/Immunofluorescence analysis of Clathrin shows staining in NCI-H460 cells. Clathrin, Heavy chain staining (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 ab2731 (1:200) over night at 4°C, washed with PBS and incubated with a DyLight-488 conjugated goat anti-mouse secondary antibody. Images were taken at 60X magnification.

  • Immunocytochemistry/Immunofluorescence analysis of Clathrin shows staining in U251 cells. Clathrin, Heavy chain staining (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 ab2731 (1:200) over night at 4°C, washed with PBS and incubated with a DyLight-488 conjugated goat anti-mouse secondary antibody. Images were taken at 60X magnification.

  • ab2731 staining human HEK 293 cells by ICC/IF.  Cells were PFA fixed and permeabilized in 0.01% Triton X-100 in PBS prior to blocking in 1% BSA for 1 hour at 22°C.  The primary antibody was diluted 1/75 and incubated with the sample for 1 hour at 22°C.  An Alexa Fluor® 405 conjugated goat anti-mouse antibody diluted 1/400 was used as the secondary.

    See Abreview

  • All lanes : Anti-Clathrin antibody [X22] (ab2731) at 1/500 dilution

    Lane 1 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 2 : Raji (Human Burkitt's lymphoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Observed band size : 180 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 240 kDa,450 kDa. We are unsure as to the identity of these extra bands.
  • ab2731 staining Clathrin - Membrane Vesicle Marker in Rat hypothalamus primary cells by ICC/IF (immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.5% TX-100 and blocked with 5% serum for 20 minutes at 25°C. Samples were incubated with primary antibody (1/500 in 1% goat serum; 0.1% TX100; PBS) for 16 hours at 4°C. An Alexa Fluor®546-conjugated Goat polyclonal to mouse IgG, dilution 1/500, was used as secondary antibody.

    See Abreview

  • ab2731 staining Clathrin in pig retinal pigment epithelium (RPE) cells by Immunocytochemistry/ Immunofluorescence. Cells were fixed with paraformaldehyde, permeabilized with 0.5% Triton ×100 and blocked with 5% serum for 20 minutes at 250C. Samples were incubated with primary antibody (1/500: in 0.1% TX100, 1% goat serum, 1X PBS) for 16 hours at 4°C. An Alexa Fluor®546-conjugated goat polyclonal to mouse IgG was used as secondary antibody at 1/500 dilution.

    See Abreview

  • ab2731 staining clathrin in Human platelets by Flow Cytometry. Platelets were isolated form Platelet rich plasma and suspended in PBS, fixed with paraformaldehyde and permeabilized with 0.1% Triton-X100 in 2% BSA for 30 minutes. The sample was incubated with the primary antibody (1/200 in PBS + 2% BSA) for 18 hours at 4°C. An Alexa Fluor®594-conjugated Goat anti-mouse IgG polyclonal (1/500) was used as the secondary antibody.
    Gating Strategy: Platelets

    See Abreview

  • Immunohistochemistry was performed on cancer biopsies of deparaffinized Human breast carcinoma tissue. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:100 with a mouse monoclonal antibody recognizing Clathrin Heavy chain ab2731 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

  • Immunohistochemistry was performed on normal biopsies of deparaffinized Human colon tissue. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:100 with a mouse monoclonal antibody recognizing Clathrin Heavy chain ab2731 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

  • Xenopus laevis cytoplasmic egg extract visualized live with primary and secondary antibody addition [red is anti-Clathrin X22 (ab2731) with goat anti-mouse Alexa Fluor 568 secondary, green is anti-HIP1R (ab77297) with goat anti-rabbit Alexa Fluor 488 secondary]. Large red structures are probably aggregates, but the small structures appear to be specific for vesicle staining.

    See Abreview

References for Anti-Clathrin antibody [X22] (ab2731)

This product has been referenced in:
  • Salinas S  et al. Disruption of the coxsackievirus and adenovirus receptor-homodimeric interaction triggers lipid microdomain- and dynamin-dependent endocytosis and lysosomal targeting. J Biol Chem 289:680-95 (2014). Mouse . Read more (PubMed: 24273169) »
  • Sullivan CS  et al. The adaptor protein GULP promotes Jedi-1-mediated phagocytosis through a clathrin-dependent mechanism. Mol Biol Cell 25:1925-36 (2014). IP . Read more (PubMed: 24743597) »

See all 25 Publications for this product

Product Wall

We have never experienced this problem with this antibody. Please note, that this antibody has not been tested in immunohistochemistry on frozen tissue sections and is therefore not covered by the abpromise guarantee.

This antibody has been u...

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I sent an proforma with the number 48311 for the Anti-Clathrin antibody [X22] - Membrane Vesicle Marker (ab2731). Please read our offer regarding the RabMab antibodies at the end of this email, it might be useful f...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample African Green Monkey Cell lysate - other (CV1 cells)
Loading amount 10 µg
Specification CV1 cells
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Mr. Nir Drayman

Verified customer

Submitted Sep 11 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry
Sample Mouse Cell (RAW 264.7)
Specification RAW 264.7
Fixative Paraformaldehyde
Permeabilization Yes - 0.1% Triton-X100 in 2% BSA for 15min
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 22°C
Username

Dr. Mahesh Shivananjappa

Verified customer

Submitted May 24 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Xenopus laevis Cell (Xenopus laevis cytoplasmic egg extract)
Specification Xenopus laevis cytoplasmic egg extract
Fixative live, no fixative
Permeabilization No
Username

Abcam user community

Verified customer

Submitted Apr 17 2012

Application Western blot
Sample Xenopus laevis Cell lysate - other (Xenopus laevis cytoplasmic egg extract)
Loading amount 40 µg
Specification Xenopus laevis cytoplasmic egg extract
Gel Running Conditions Reduced Denaturing (10%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Apr 17 2012



Nous avons dans notre catalogue des anticorps marqueurs des voies d'internalisation tels que les anti-Clathrin ab2731 (http://www.abcam.com/ab2731) et ab59710 (http://www.abcam.com/ab59710). Malheureusement nous n'avons pas ces anticorps coup...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (Platelets)
Loading amount 20 µg
Specification Platelets
Treatment ADP for 30 min
Gel Running Conditions Reduced Denaturing (4-20%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Dr. Mahesh Shivananjappa

Verified customer

Submitted Nov 24 2011

Thank you for your reply. All Abreviews we have are from previous batches of this antibody. This antibody is very popular and sells fast. Therefore even the experiment shown in the latest Abreview was performed with a different batch. I...

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