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

Reviews (17)Q&A (8)References (64)

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

  • 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.

  • 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

製品の特性

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
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

画像

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

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

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

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

    See Abreview

  • Immunocytochemistry/ Immunofluorescence - Anti-Clathrin antibody [X22] (ab2731)
    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.

    See Abreview

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

    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 (64)

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 64 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
  • 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
  • Tse A  et al. Strong stimulation triggers full fusion exocytosis and very slow endocytosis of the small dense core granules in carotid glomus cells. J Neurogenet 32:267-278 (2018). PubMed: 30484390
View all Publications for this product

レビューと Q&A

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1-10 of 25 Abreviews or Q&A

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.

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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.

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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,

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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

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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,

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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.

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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
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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
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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
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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
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Abcam user community

Verified customer

投稿 Apr 17 2012

Question

infos sur des marqueurs fluo des voies d'internalisation.

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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.

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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
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Dr. Mahesh Shivananjappa

Verified customer

投稿 Nov 24 2011

Question

thank you for your reply. Our customer would like to know if you have any abreview for both or for just one these batches about WB analysis. If so, please send me all the information you have.   Thanks in advance Kind regards  

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Answer

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 am sorry I cannot provide you with this information, but I would like to reassure you and your customer that at Abcam our products are covered by the Abpromise. Our Abpromise® ensures that you can trust our products to perform as stated on the datasheets, or we will offer a replacement, credit, or refund. The Abcam Abpromise® guarantee: • 100% Scientific and Customer Support for any product you buy from Abcam or one of our authorised distributors. • We guarantee our products work in the tested species and applications stated on the datasheet. • We will replace or refund products not performing as stated on the datasheet if reported within 6 months of purchase. • We investigate any quality concerns raised by customers to ensure our catalog contains products that perform to the highest standards. Please note these conditions to our Abpromise®: • Protocol information must be provided in order for the claim to be validated. • Antibodies tested in recombinant samples only are not guaranteed for use on endogenous samples. • “Predicted to react” information on the datasheets is provided for reference only and these species are not guaranteed. • Fast Track antibodies are covered for ELISA against the immunizing peptide only. I hope this information is helpful, but please do not hesitate to contact us with further questions.

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1-10 of 25 Abreviews or Q&A

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