Anti-Clathrin heavy chain 抗体 [X22] (ab2731)

Reviews (18)Q&A (8)References (66)
Immunocytochemistry - Anti-Clathrin heavy chain antibody [X22] (ab2731)
  • Western blot - Anti-Clathrin heavy chain antibody [X22] (ab2731)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Clathrin heavy chain antibody [X22] (ab2731)
  • Immunocytochemistry - Anti-Clathrin heavy chain antibody [X22] (ab2731)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Clathrin heavy chain antibody [X22] (ab2731)
  • Immunocytochemistry - Anti-Clathrin heavy chain antibody [X22] (ab2731)
  • Immunocytochemistry - Anti-Clathrin heavy chain antibody [X22] (ab2731)
  • Western blot - Anti-Clathrin heavy chain antibody [X22] (ab2731)

Key features and details

  • Mouse monoclonal [X22] to Clathrin heavy chain
  • Suitable for: WB, IHC-P, ICC
  • Reacts with: Human, Xenopus laevis
  • Isotype: IgG1

リコンビナント抗体で、ロット間での高い再現性を実現

Product image
Anti-Clathrin heavy chain antibody [EPR12235(B)] (ab172958)
  • 異なるロット間での安定した再現性
  • 容易なスケールアップ
  • 評価試験による特異性の確認済み
  • 倫理基準に準拠 - アニマル・フリーの生産

製品の概要

  • 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: WB, IHC-P, ICCmore details

  • 種交差性

    交差種: Human, Xenopus laevis

  • 免疫原

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

  • 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.

  • 特記事項

    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

製品の特性

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab2731の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
WB (5)
1/100 - 1/500. Detects a band of approximately 180 kDa.
IHC-P
1/100.
ICC (2)
1/1000.
特記事項
WB
1/100 - 1/500. Detects a band of approximately 180 kDa.
IHC-P
1/100.
ICC
1/1000.

ターゲット情報

  • 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

  • 参照データベース

    • 別名

      • CHC17 antibody
      • Clathrin heavy chain antibody
      • Clathrin heavy chain 1 antibody
      • Clathrin heavy chain 2 antibody
      • Clathrin heavy chain like 1 antibody
      • Clathrin heavy chain on chromosome 17 antibody
      • Clathrin heavy polypeptide antibody
      • CLH 17 antibody
      • CLH 22 antibody
      • CLH-17 antibody
      • CLH1_HUMAN antibody
      • CLH17 antibody
      • CLH22 antibody
      • CLTC antibody
      • CLTCL antibody
      • CLTCL1 antibody
      • CLTCL2 antibody
      • CLTD antibody
      • Hc antibody
      • KIAA0034 antibody
      see all

    画像

    • Immunocytochemistry - Anti-Clathrin heavy chain antibody [X22] (ab2731)
      Immunocytochemistry - 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.

    • Western blot - Anti-Clathrin heavy chain antibody [X22] (ab2731)
      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)
      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 - Anti-Clathrin heavy chain antibody [X22] (ab2731)
      Immunocytochemistry - 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.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Clathrin heavy chain antibody [X22] (ab2731)
      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 - Anti-Clathrin heavy chain antibody [X22] (ab2731)
      Immunocytochemistry - 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.

    • Immunocytochemistry - Anti-Clathrin heavy chain antibody [X22] (ab2731)
      Immunocytochemistry - 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.

      See Abreview

    • Western blot - Anti-Clathrin heavy chain antibody [X22] (ab2731)
      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.

    参考文献 (66)

    ab2731 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    ab2731 は 66 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
    • Fay NC  et al. A Novel Fusion of IL-10 Engineered to Traffic across Intestinal Epithelium to Treat Colitis. J Immunol 205:3191-3204 (2020). PubMed: 33148717
    • 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
    View all Publications for this product

    レビューと Q&A

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    レビューを送る 質問を送る

    1-10 of 26 Abreviews or Q&A

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Clathrin heavy chain antibody [X22]

     Good
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa)
    Permeabilization
    Yes - 0.5% Triton X-100 in PBS, 5min, RT
    Specification
    HeLa
    Fixative
    Paraformaldehyde
    Read More

    Carlos Pardo-Pastor

    Verified customer

    投稿 Apr 13 2021

    Question

    We are doing some IF research on frozen rat brains and have always been very satisfied when purchasing antibodies from your company. However the Clathrin antibody is posing some difficulties that we haven't encountered before. When we look at the stained slides the we can see the antibody but it seems to almost have diffused out in the mounting medium as well as we find it all over the slides and not only on the tissue. Is this a known problem with this antibody and can it be sorted out by changing something? The frozen slides become fixated in 4% PFA for 1 hour at first before pre-blocking. The pre-blocking step is performed with 2,5% horseserum in PBS for one hour.

    Read More
    Answer

    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 used on paraffin slides at a dilution of 1:100, therefore, I would suggest using a similar concentration for IHC-Fr. Since the antibody is diluted down, it is more likely that the problem is due to the dilution buffer than the antibody itself. We have in the past got very successful results using 3% BSA-PBS as the diluent so I would suggest trying this.

    Read More

    Question

    Inquiry: To whom it may concern, I am writing you to recorrect my previously sent email address. This is the proper email for which I am waiting your answer. I I would like to inquire about Anti-Clathrin antibody [X22] - Membrane Vesicle Marker (ab2731). Could you ship it to Hungary? Would you be so kind to send a preformal invoice to the above email address with the price of product and shipping? Thank you for your help in advance. Kind regards,

    Read More
    Answer

    Thank you for contacting us.

    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 for you.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Free Rabbit monoclonal antibody with any purchase of a primary antibody, while stocks last! Quote “RABMAB-XBSMG” in your next primary antibody order. For more information, visit the following link: https://www.abcam.com/index.html?pageconfig=resource&rid=15447

    Read More

    Question

    Inquiry: Dear Sir/Madam, I am planning a large scale experiment using one for the following antibodies but I need to test a small amount from each before ordering large scale. I was wondering if you supply sample free stocks I could test? Abcam ab6323 Anti-beta COP antibody [maD] (ab6323) Abcam ab99552 Anti-SEC23 antibody (ab99552) Abcam ab2731 Anti-Clathrin antibody [X22] - Membrane Vesicle Marker (ab2731) Thanks,

    Read More
    Answer

    Thank you for contacting us. Because we carry over 95,000 products, it isn't feasible for us to keep small sample sizes of our products.

    We are happy to reassure our customers that all of our products are covered by our Abpromise, which guarantees that the product will work in the applications and species specified on the datasheet, or we will offer a replacement, credit, or refund within 6 months of purchase.

    If the product is to be used in an untested species or application, you may be eligible for our testing discount program if the antibody has not yet been purchased. Please contact our Scientific Support team by replying to this email prior to purchase for more information.

    Otherwise, we like to encourage all of our customers to submit an Abreview via the online product datasheet. We always appreciate customer feedback, whether positive or negative, and we make all product information available to researchers. Plus, each Abreview earns Abpoints that can be used for discounts on future purchases or rewards such as Amazon.com gift certificates.

    To find out more about our Abreview system, please see the following link:

    https://www.abcam.com/abreviews



    I hope this information is helpful. Please do not hesitate to contact us again with any other questions.

    Read More

    Western blot abreview for Anti-Clathrin antibody [X22] - Membrane Vesicle Marker

     Excellent
    Abreviews
    abreview image
    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
    Read More

    Mr. Nir Drayman

    Verified customer

    投稿 Sep 11 2012

    Immunocytochemistry abreview for Anti-Clathrin antibody [X22] - Membrane Vesicle Marker

     Excellent
    Abreviews
    abreview image
    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
    Read More

    Dr. Mahesh Shivananjappa

    Verified customer

    投稿 May 24 2012

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Clathrin antibody [X22] - Membrane Vesicle Marker

     Good
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Xenopus laevis Cell (Xenopus laevis cytoplasmic egg extract)
    Specification
    Xenopus laevis cytoplasmic egg extract
    Fixative
    live, no fixative
    Permeabilization
    No
    Read More

    Abcam user community

    Verified customer

    投稿 Apr 17 2012

    Western blot abreview for Anti-Clathrin antibody [X22] - Membrane Vesicle Marker

     Poor
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    投稿 Apr 17 2012

    Question

    infos sur des marqueurs fluo des voies d'internalisation.

    Read More
    Answer



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

    Nous avons cependant des kits de conjugaison, les kits EasyLink, qui permettent de coupler rapidement et facilement les anticorps primaire. Pour plus d'informations sur ces kits je vous invite à visiter la page suivante : https://www.abcam.com/easylink.

    Pour les voies d'internalisation je vous suggère le lien suivant : https://www.abcam.com/index.html?pageconfig=resource&rid=10236&pid=14.

    J'espère que ces informations vous seront utiles. N'hésitez pas à nous contacter de nouveau si vous avez d'autres questions.

    Read More

    Western blot abreview for Anti-Clathrin antibody [X22] - Membrane Vesicle Marker

     Average
    Abreviews
    abreview image
    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
    Read More

    Dr. Mahesh Shivananjappa

    Verified customer

    投稿 Nov 24 2011

    1-10 of 26 Abreviews or Q&A

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