Anti-Clathrin heavy chain 抗体 [X22] (ab2731)
Key features and details
- Mouse monoclonal [X22] to Clathrin heavy chain
- Suitable for: ICC/IF, WB, IHC-P
- Reacts with: Mouse, Rat, Cow, Dog, Human, Bird
- Isotype: IgG1
製品の概要
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製品名
Anti-Clathrin heavy chain antibody [X22]
Clathrin heavy chain 一次抗体 製品一覧 -
製品の詳細
Mouse monoclonal [X22] to Clathrin heavy chain -
由来種
Mouse -
特異性
Detects clathrin heavy chain. -
アプリケーション
適用あり: ICC/IF, WB, IHC-Pmore details -
種交差性
交差種: Mouse, Rat, Cow, Dog, Human, Bird -
免疫原
Full length native protein (purified) corresponding to Human Clathrin heavy chain. Purified human brain clathrin heavy chain.
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エピトープ
Electron microscopy and proteolysis mapping demonstrate that binding occurs towards the central hub of the triskelion, N-terminal to the light chain binding regions. -
ポジティブ・コントロール
- bovine brain extract
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特記事項
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
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.
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, as well as customer reviews and Q&As.
製品の特性
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製品の状態
Liquid -
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
Constituent: PBS -
Concentration information loading...
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精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
X22 -
アイソタイプ
IgG1 -
研究分野
関連製品
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Recombinant Protein
アプリケーション
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.
アプリケーション | Abreviews | 特記事項 |
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ICC/IF | 1/1000. | |
WB | 1/500. Detects a band of approximately 180 kDa. | |
IHC-P | 1/100. |
ターゲット情報
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機能
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. -
配列類似性
Belongs to the clathrin heavy chain family. -
細胞内局在
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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参照データベース
- Entrez Gene: 281080 Cow
- Entrez Gene: 480578 Dog
- Entrez Gene: 1213 Human
- Entrez Gene: 67300 Mouse
- Entrez Gene: 54241 Rat
- Omim: 118955 Human
- SwissProt: P49951 Cow
- SwissProt: P53675 Human
see all -
別名
- CHC17 antibody
- Clathrin heavy chain antibody
- Clathrin heavy chain 1 antibody
see all
画像
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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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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)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.
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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)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.
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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)This image is courtesy of an Abreview submitted by Dr Vladimir Milenkovic
ab2731 staining Clathrin heavy chain - 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.
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Immunocytochemistry/ Immunofluorescence - Anti-Clathrin antibody [X22] (ab2731)This image is a courtesy of Vladimir Milenkovic
ab2731 staining Clathrin heavy chain 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.
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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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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.
プロトコール
データシートおよび資料
参考文献 (65)
ab2731 は 65 報の論文で使用されています。
- Ren F et al. NRSN2 promotes breast cancer metastasis by activating PI3K/AKT/mTOR and NF-?B signaling pathways. Oncol Lett 19:813-823 (2020). PubMed: 31885716
- Taverner A et al. Cholix protein domain I functions as a carrier element for efficient apical to basal epithelial transcytosis. Tissue Barriers 8:1710429 (2020). PubMed: 31928299
- Moshkanbaryans L et al. The interaction of assembly protein AP180 and clathrin is inhibited by multi-site phospho-mimetics. Neurochem Int 129:104474 (2019). PubMed: 31129113
- Cohen EAK et al. Resolution limit of image analysis algorithms. Nat Commun 10:793 (2019). PubMed: 30770826
- Berndt-Paetz M et al. IC/BPS-associated alterations of M2 and M3 muscarinic acetylcholine receptor trafficking in human detrusor. Neurourol Urodyn 38:1818-1827 (2019). PubMed: 31301091