Anti-Clathrin heavy chain antibody [X22] (ab2731)
![Immunocytochemistry/ Immunofluorescence - Anti-Clathrin heavy chain antibody [X22] (ab2731)](https://www.abcam.com/ps/products/2/ab2731/Images/ab2731-218980-anti-clathrin-antibody-x22-immunofluorescence.jpg "Immunocytochemistry/ Immunofluorescence - Anti-Clathrin heavy chain antibody [X22] (ab2731)")
Key features and details
- Mouse monoclonal [X22] to Clathrin heavy chain
- Suitable for: WB, IHC-P, ICC/IF
- Reacts with: Human, Xenopus laevis
- Isotype: IgG1
Overview
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Product name
Anti-Clathrin heavy chain antibody [X22]
See all Clathrin heavy chain primary antibodies -
Description
Mouse monoclonal [X22] to Clathrin heavy chain -
Host species
Mouse -
Tested applications
Suitable for: WB, IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Human, Xenopus laevis -
Immunogen
Full length native protein (purified) corresponding to Human Clathrin heavy chain. Purified human brain clathrin heavy chain.
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Epitope
Electron microscopy and proteolysis mapping demonstrate that binding occurs towards the central hub of the triskelion, N-terminal to the light chain binding regions. -
General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
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. -
Storage buffer
Preservative: 0.05% Sodium azide
Constituent: PBS -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
X22 -
Isotype
IgG1 -
Research areas
Associated products
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
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 |
|---|---|---|
| WB | (5) |
1/100 - 1/500. Detects a band of approximately 180 kDa.
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| IHC-P |
1/100.
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| ICC/IF | (11) |
1/1000.
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| Notes |
|---|
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WB
1/100 - 1/500. Detects a band of approximately 180 kDa. |
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IHC-P
1/100. |
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ICC/IF
1/1000. |
Target
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Function
Clathrin is the major protein of the polyhedral coat of coated pits and vesicles. Two different adapter protein complexes link the clathrin lattice either to the plasma membrane or to the trans-Golgi network. -
Sequence similarities
Belongs to the clathrin heavy chain family. -
Cellular localization
Cytoplasmic vesicle membrane. Membrane > coated pit. Melanosome. Cytoplasmic face of coated pits and vesicles. Identified by mass spectrometry in melanosome fractions from stage I to stage IV. - Information by UniProt
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Database links
- Entrez Gene: 1213 Human
- Omim: 118955 Human
- SwissProt: P53675 Human
- SwissProt: Q00610 Human
- Unigene: 491351 Human
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Alternative names
- CHC17 antibody
- Clathrin heavy chain antibody
- Clathrin heavy chain 1 antibody
see all
Images
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Immunocytochemistry/ Immunofluorescence - Anti-Clathrin heavy chain antibody [X22] (ab2731)
Immunocytochemistry/Immunofluorescence analysis of Clathrin heavy chain 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.
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![Western blot - Anti-Clathrin heavy chain antibody [X22] (ab2731)](https://www.abcam.com/ps/products/2/ab2731/Images/ab2731-275742-anti-clathrin-antibody-x22-western-blot.png)
Western blot - Anti-Clathrin heavy chain antibody [X22] (ab2731)Anti-Clathrin heavy chain antibody [X22] (ab2731) at 1/300 dilution + Human brain lysates at 25 µg
Secondary
HRP-conjugated goat anti-mouse IgG + IgM (H+L)
Developed using the ECL technique. -
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Clathrin heavy chain antibody [X22] (ab2731)](https://www.abcam.com/ps/products/2/ab2731/Images/ab2731-6268-anti-clathrin-antibody-x22-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Clathrin heavy chain antibody [X22] (ab2731)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. -
![Immunocytochemistry/ Immunofluorescence - Anti-Clathrin heavy chain antibody [X22] (ab2731)](https://www.abcam.com/ps/products/2/ab2731/Images/ab2731-218978-anti-clathrin-antibody-x22-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-Clathrin heavy chain antibody [X22] (ab2731)Immunocytochemistry/Immunofluorescence analysis of Clathrin heavy chain 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.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Clathrin heavy chain antibody [X22] (ab2731)](https://www.abcam.com/ps/products/2/ab2731/Images/ab2731-6269-anti-clathrin-antibody-x22-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Clathrin heavy chain antibody [X22] (ab2731)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. -
![Immunocytochemistry/ Immunofluorescence - Anti-Clathrin heavy chain antibody [X22] (ab2731)](https://www.abcam.com/ps/products/2/ab2731/Images/ab2731-218977-anti-clathrin-antibody-x22-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-Clathrin heavy chain antibody [X22] (ab2731)Immunocytochemistry/Immunofluorescence analysis of Clathrin heavy chain 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.
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![Immunocytochemistry/ Immunofluorescence - Anti-Clathrin heavy chain antibody [X22] (ab2731)](https://www.abcam.com/ps/products/2/ab2731/Images/ab2731-6267-anti-clathrin-antibody-x22-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-Clathrin heavy chain antibody [X22] (ab2731)Xenopus laevis cytoplasmic egg extract visualized live with primary and secondary antibody addition [red is anti-Clathrin heavy chain 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.
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![Western blot - Anti-Clathrin heavy chain antibody [X22] (ab2731)](https://www.abcam.com/ps/products/2/ab2731/Images/ab2731-6273-anti-clathrin-antibody-x22-western-blot.jpg)
Western blot - Anti-Clathrin heavy chain antibody [X22] (ab2731)All lanes : Anti-Clathrin heavy chain 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
All lanes : 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.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (80)
ab2731 has been referenced in 80 publications.
- Raykova D et al. A method for Boolean analysis of protein interactions at a molecular level. Nat Commun 13:4755 (2022). PubMed: 35963857
- Yang C et al. Actin polymerization promotes invagination of flat clathrin-coated lattices in mammalian cells by pushing at lattice edges. Nat Commun 13:6127 (2022). PubMed: 36253374
- Pasquettaz R et al. Peculiar protrusions along tanycyte processes face diverse neural and nonneural cell types in the hypothalamic parenchyma. J Comp Neurol 529:553-575 (2021). PubMed: 32515035
- Hernaiz-Llorens M et al. Growth cone repulsion to Netrin-1 depends on lipid raft microdomains enriched in UNC5 receptors. Cell Mol Life Sci 78:2797-2820 (2021). PubMed: 33095273
- Mau A et al. Fast widefield scan provides tunable and uniform illumination optimizing super-resolution microscopy on large fields. Nat Commun 12:3077 (2021). PubMed: 34031402