Anti-Nuclear Pore Complex Proteins antibody [Mab414] - ChIP Grade (ab24609)

Mouse monoclonal Nuclear Pore Complex Proteins antibody [Mab414]. Validated in WB, IP, IHC, ChIP, EM, ICC/IF and tested in Mouse, Rat, Cat, Human, Saccharomyces cerevisiae, Xenopus laevis and more.

Overview

  • Product name
    Anti-Nuclear Pore Complex Proteins antibody [Mab414] - ChIP Grade
    See all Nuclear Pore Complex Proteins primary antibodies
  • Description
    Mouse monoclonal [Mab414] to Nuclear Pore Complex Proteins - ChIP Grade
  • Host species
    Mouse
  • Specificity
    ab24609 reacts with Nuclear Pore Complex (NPC) Proteins. Aris et al ( J. Cell Biol. 108:2059-2067, 1989) report "In rat liver, this monoclonal antibody, mAb 414, binds to nuclear pore complex proteins, including one of molecular weight 62,000 (Davis, L. I., and G. Blobel. 1987. Proc. Natl. Acad. Sci. USA. 84:7552-7556). In yeast, mAb 414 cross reacts by immunoblotting with two proteins that have apparent molecular weights of 110,000 and 95,000, and are termed p110 and p95, respectively. Examination of subcellular fractions by immunoblotting shows that both p110 and p95 are located exclusively in the nuclear fraction. The mAb 414 immunoprecipitates several proteins from a crude yeast cell extract, including p110, p95, and a approximately 55-kD protein. Immunoprecipitation from subcellular fractions yields only p110 and p95 from purified nuclei, whereas the approximately 55-kD protein is immunoprecipitated from the soluble fraction. Digestion of purified nuclei with DNase to produce nuclear envelopes releases some of p110, but the majority of p110 is solubilized only after treatment of envelopes with 1 M NaCl. Immunofluorescence localization using yeast cells and isolated nuclei shows a punctate and patchy staining pattern of the nucleus."
  • Tested applications
    Suitable for: ChIP, WB, ICC/IF, IP, Electron Microscopy, IHC-Frmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Cat, Human, Saccharomyces cerevisiae, Xenopus laevis, Caenorhabditis elegans, Drosophila melanogaster, Zebrafish
    Predicted to work with: Vertebrata
  • Immunogen

    Nuclear pore complex mixture.

  • Epitope
    ab24609 recognizes the conserved domain FXFG repeats in nucleaporins like the p62, p152, p90.
  • Positive control
    • Raw, HEK 293 cell lysate (see Abreview), rat liver lysate (see Aris reference)
  • General notes


    This is a reliable general purpose monoclonal antibody which recognizes a related family of NPC proteins. This antibody is ideal for studying the morphology and composition of the nucleus and nuclear envelope. It is also useful in studying changes in the nuclear structure during mitosis and meiosis.

Properties

  • 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: 0.03% Thimerosal (merthiolate)
    Constituent: PBS
  • Concentration information loading...
  • Purity
    Ascites
  • Purification notes
    Purified from ascites.
  • Primary antibody notes
    This is a reliable general purpose monoclonal antibody which recognizes a related family of NPC proteins. This antibody is ideal for studying the morphology and composition of the nucleus and nuclear envelope. It is also useful in studying changes in the nuclear structure during mitosis and meiosis.
  • Clonality
    Monoclonal
  • Clone number
    Mab414
  • Isotype
    IgG1
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab24609 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
ChIP Use at an assay dependent concentration. PubMed: 20419146
WB 1/5000. Predicted molecular weight: 62 kDa. Suitable also in non reduced western blotting conditions (see Abreview). Nuclear extraction may be necessary (protocol detailed in Aris et al, http://www.jcb.org/cgi/reprint/108/6/2059).
ICC/IF 1/5000. Fix cells with 4% paraformaldehyde in NWB (200 mM sucrose, 15 mM Hepes, pH 7.4, 50 mM NaCl, 2.5 mM MgCl2, and 1 mM DTT). Permeabilise with 0.1% NP-40 or 0.1% Triton X-100 in PBS for 2 min. (see Lopez-Soler reference); different customer have used this antibody at different dilutions for ICC/IF (see images below). We recommend that optimal working dilutions are determined by each customer.
IP 1/5000. Nuclear extraction may be necessary (protocol detailed in Aris et al, http://www.jcb.org/cgi/reprint/108/6/2059).
Electron Microscopy 1/5000.
IHC-Fr Use at an assay dependent concentration. 4% paraformaldehyde fixed tissue cut on a cryostat (see Kimura reference).

Target

Images

  • ab24609 staining nuclear pore complex proteins in Human chronic myelogenous leukemia cells from bone marrow cells. Cells were fixed with 4% paraformaldehyde in DPBS buffer for 20 mins and permeabilised with 0.1% Triton X-100 for 20 min at room temperature. Cells were blocked with 2% normal goat serum in DPBS with 1% BSA and washed with 0.1% Tween 20.

  • ab24609 (1/200) staininig nuclear pore complex proteins in human RPE-1 cells (green). Cells were fixed in paraformaldehyde, permeabilised with Triton X100 and counterstained with DAPI in order to highlight the nucleus (blue). Please refer to abreview for further experimental details.

    See Abreview

  • ab24609 (1/500) staining nuclear pore complex proteins in human Hela Cells (green). Cells were fixed with Paraformaldehyde/Triton X-100 (10 min in PTEMF buffer (20mM PIPES, 1mM MgCl2, 10mM EGTA, 4% PFA) /0.2% Triton-X100 at room T°C) or Methanol (6 min in Methanol -20 °C , followed by 3 washes in 1x PBS) and counterstained with Dapi in order to highligh the nucleus (blue).
  • ab24609 (1/500) staining nuclear pore complex proteins in murine NIH/3T3 Cells (green). Cells were fixed with Paraformaldehyde/Triton X-100 (10 min in PTEMF buffer (20mM PIPES, 1mM MgCl2, 10mM EGTA, 4% PFA) /0.2% Triton-X100 at room T°C) or Methanol (6 min in Methanol -20 °C , followed by 3 washes in 1x PBS) and counterstained with Dapi in order to highligh the nucleus (blue).

References

This product has been referenced in:
  • Bejarano DA  et al. HIV-1 nuclear import in macrophages is regulated by CPSF6-capsid interactions at the nuclear pore complex. Elife 8:N/A (2019). Read more (PubMed: 30672737) »
  • Pereira C  et al. Self-Assembly of the RZZ Complex into Filaments Drives Kinetochore Expansion in the Absence of Microtubule Attachment. Curr Biol 28:3408-3421.e8 (2018). Read more (PubMed: 30415699) »
See all 74 Publications for this product

Customer reviews and Q&As

1-10 of 12 Q&A

Question
Answer

The staining pattern of this antibody should be nuclear or nuclear membrane.

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Answer

Thank you for contacting us.



While this product is predicted to react with all vertebrates, we have not yet tested this with cow. For UNTESTED species and/or applications, we have established a testing discount program. Here is a brief description of how it works:



The testing discount program is for customers who like to use an antibody/protein on an untested species/application. You would purchase the antibody at full price, test it and submit an Abreview with your data (positive or negative). On your next order you will receive a discount for ONE antibody at the full price (100%) of the antibody you have tested. The terms and conditions applicable to this offer can be found here: https://www.abcam.com/collaborationdiscount.





I hope this information is helpful. Please let me know if you would like to participate in this program and test in cow. Also, please do not hesitate to contact us if you need any more advice or information.



Use our products? Submit an Abreview. Earn rewards!

https://www.abcam.com/abreviews

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Answer

I am very pleased to hear you would like to accept our offer and test one or both of these products in blocking experiements. Each code is associated with a paricular antibody so be sure tho reference the correct code when ordering and submitting the Abreview. Each will give you: 1 free PRIMARY ANTIBODY before the expiration date. To redeem this offer, please submit an Abreview and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. Please remember that submission of the Abreview is sufficient for the discount code to become active. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews. Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code. Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research. The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

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Answer

Thank you very much for your interest in our Nuclear Pore Complex Proteins antibodies. I have spoken to the labs about these product and have found that these are indeed the same clone detailed inin the papers by Davis & Blobel as we spoke of.

We do not currently have a testing data using our Nuclear Pore Complex Proteins antibodies, ab24609 or ab50008in [blocking experiments. Therefore, I can offer a discount off a future purchase if you buy one of these products now, use it and submit an image of your results to us in the form of an Abreview. The discount would be to the value of 1 free PRIMARY ANTIBODY.

It doesn’t matter whether the result is positive or negative, we would just really like to receive your feedback. Of course, if the results are negative you will be covered by our Abpromise guarantee and eligible for a refund or replacement.

If you are interested in this offer, please follow these steps:

1. Reply to this e-mail to let me know that you would like to proceed to test and submit an Abreview in a blocking experiment. I will then send a discount code. This code must be issued before purchasing so please wait for my reply before ordering.

2. Purchase either by phone, fax, or online (www.abcam.com).

3. Use it in your blocking experiment.

4. Submit an Abreview of the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.

5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any PRIMARY ANTIBODY ordered and the discount code is valid for 4 months after issue.
Please remember that submission of the Abreview is sufficient for the discount code to become active.

We are always pleased to obtain feedback about our products and any information is greatly appreciated!

Please let me know which product you would like to test and if you have any questions about this offer and I would be happy to help you further.

The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

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Answer

Thank you very much for your interest in ab24609.

To our knowledge, ab24609 has not been tested in IHC-P. Therefore, I can offer a discount off a future purchase if you buy ab24609 now, test it inIHC-P and submit feedback to us in the form of an Abreview. It doesn't matter whether the Abreview is positive or negative, we would just really like to receive your feedback. The discount would be to the value of 1 free primary antibody.

If you are interested in this offer, please follow these steps:

1. Reply to this e-mail to let me know that you would like to proceed and test ab24609 in IHC-P. I will then send a discount code. This code must be issued before purchasing ab24609 so please wait for my reply before ordering.

2. Purchase ab24609 either by phone, fax, or online (www.abcam.com).

3. Test it in IHC-P.


4. Let us know the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews.

5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for anyprimary antibodyordered and the discount code is valid for 4 months after issue.

We are always pleased to obtain feedback about our products and any information is greatly appreciated! Even if ab24609 turns out to be unsuitable for IHC-P, you will still receive the discount on your next purchase after your Abreview has been submitted.

Please let me know if you have any questions about this offer and I would be happy to help you further.

The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

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Answer

Thank you for contacting us. The review has been updated. The updates will appear on the website soon. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.  

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Answer

Thank you for contacting Abcam. I am sorry for the confusion over the alternative names for ab24609, but I think what has happened is that because ab24609 recognizes multiple Nuclear Pore Complex Proteins and Nup107 being a nuclear pore complex protein, then Nup107 may have been added in error. I will forward the information you sent to out relevant department so they can correct any mistakes. If there is anything else I can help you with, please let me know.

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Answer

DISCOUNT CODE: XXXXXXXXXXX Expiration date: 04/02/2012 I am very pleased to hear you would like to accept our offer and test ab24609 in Feline species. This code will give you 1 free PRIMARY ANTIBODY before the expiration date. To redeem this offer, please submit an Abreview for Feline and include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews. Remember, we publish both positive and negative Abreviews on our datasheets so please submit the results of your tests. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code. Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research. The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

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Answer

Thank you for contacting us. Because we carry over 70,000 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. As far as we are aware, ab24609 has never been tested with feline cells or tissues. All tested applications are specified on our datasheets, and these are updated as soon as any new information is brought to our attention. I can however offer a discount off a future purchase if you buy ab24609 now, use it in Feline species sample and submit an image of your results to us in the form of an Abreview. The discount would be to the value of 1 free PRIMARY ANTIBODY. It doesn’t matter whether the result is positive or negative, we would just really like to receive your feedback. Of course, if the results are negative you will be covered by our Abpromise guarantee and eligible for a refund or replacement. If you are interested in this offer, please follow these steps: 1. Reply to this e-mail to let me know that you would like to proceed and send an image of ab24609 in Feline species. I will then send a discount code. This code must be issued before purchasing ab24609 so please wait for my reply before ordering. 2. Purchase ab24609 either by phone, fax, or online (www.abcam.com). 3. Use it in Feline species samples. 4. Send us an image of the results, positive or negative, using our Abreview system (this will take about 10 minutes and images are great if you have them!). To find out how to submit an Abreview, please visit: https://www.abcam.com/abreviews. 5. After the review is submitted to us, the discount code becomes active. Simply place your new order by phone, fax, or on the web and mention the discount code. The discount can be redeemed for any PRIMARY ANTIBODY ordered and the discount code is valid for 4 months after issue. We are always pleased to obtain feedback about our products and any information is greatly appreciated! Please let me know if you have any questions about this offer and I would be happy to help you further. The Terms and Conditions of this offer can be found at: www.abcam.com/collaborationdiscount.

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1-10 of 12 Q&A

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