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Anti-Mitotic proteins 抗体 [MPM-2] (ab14581)

Reviews (1)Q&A (5)References (11)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mitotic proteins antibody [MPM-2] (ab14581)
  • Immunocytochemistry - Anti-Mitotic proteins antibody [MPM-2] (ab14581)
  • Flow Cytometry (Intracellular) - Anti-Mitotic proteins antibody [MPM-2] (ab14581)
  • Western blot - Anti-Mitotic proteins antibody [MPM-2] (ab14581)
  • Western blot - Anti-Mitotic proteins antibody [MPM-2] (ab14581)
  • Immunocytochemistry - Anti-Mitotic proteins antibody [MPM-2] (ab14581)

Key features and details

  • Mouse monoclonal [MPM-2] to Mitotic proteins
  • Suitable for: IP, ICC/IF, ICC, ELISA, IHC-P, WB, Flow Cyt (Intra)
  • Reacts with: Human
  • Isotype: IgG1

製品の概要

  • 製品名

    Anti-Mitotic proteins antibody [MPM-2]

  • 製品の詳細

    Mouse monoclonal [MPM-2] to Mitotic proteins

  • 由来種

    Mouse

  • 特異性

    Recognizes a phosphorylated epitope (S/T)P found in phosphoproteins such as MAP2, HSP70, cdc25 and DNA topoisomerase IIa, most of which are phosphorylated at the onset of mitosis. The number of phosphoproteins recognized by MPM-2 varies from species to species and with the cell type. The clone number has been updated from (0.T.181) to (MPM-2) both clone numbers name the same antibody clone.

  • アプリケーション

    適用あり: IP, ICC/IF, ICC, ELISA, IHC-P, WB, Flow Cyt (Intra)more details

  • 種交差性

    交差種: Human
    交差が予測される動物種: a wide range of other species

  • 免疫原

    Tissue, cells or virus corresponding to Human Mitotic proteins.

  • ポジティブ･コントロール

    • WB: Colcemid treated HeLa cell lysate. ICC/IF: Saos-2 cells. IHC-P: human normal skin tissue Flow Cyt (Intra): HeLa cells.

  • 特記事項

    For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

    This product was changed from ascites to tissue culture supernatant on 11-June-19. Please note that the dilutions may need to be adjusted accordingly. If you have any questions, please do not hesitate to contact our scientific support team.

    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ab14581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
IP
Use at an assay dependent concentration.
ICC/IF (1)
Use at an assay dependent concentration.
ICC
Use at an assay dependent concentration.
ELISA
Use at an assay dependent concentration.

This antibody was used in a non-radioactive, dissociation-enhanced time resolved fluoroimmunoassa. Active enzymes were used to phosphorylate substrate peptides. The phosphorylated peptides were detected using MPM2 in conjunction with a europium (Eu3+) labeled secondary antibody. The phosphorylated peptides detected include: S1000-50 Serine Kinase Substrate Peptide (Biotin) (RRRAPLSP-N-Biotin) and S1000-80 Serine/Threonine Kinase Substrate Peptide, Recombinant, Human (Biotin).
IHC-P
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Predicted molecular weight: 92 kDa.
Flow Cyt (Intra)
Use at an assay dependent concentration.
特記事項
IP
Use at an assay dependent concentration.
ICC/IF
Use at an assay dependent concentration.
ICC
Use at an assay dependent concentration.
ELISA
Use at an assay dependent concentration.

This antibody was used in a non-radioactive, dissociation-enhanced time resolved fluoroimmunoassa. Active enzymes were used to phosphorylate substrate peptides. The phosphorylated peptides were detected using MPM2 in conjunction with a europium (Eu3+) labeled secondary antibody. The phosphorylated peptides detected include: S1000-50 Serine Kinase Substrate Peptide (Biotin) (RRRAPLSP-N-Biotin) and S1000-80 Serine/Threonine Kinase Substrate Peptide, Recombinant, Human (Biotin).
IHC-P
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Predicted molecular weight: 92 kDa.
Flow Cyt (Intra)
Use at an assay dependent concentration.

ターゲット情報

  • 細胞内局在

    The phosphoproteins have been found in the cytoplasm of mitotic cells and near centrosomes, kinetochores, and midbodies of the microtubule organizing centers.

画像

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mitotic proteins antibody [MPM-2] (ab14581)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mitotic proteins antibody [MPM-2] (ab14581)

    Ab14581 staining human normal skin. Staining is localised to the nucleus.
    Left panel: with primary antibody at 2 ug/ml. Right panel: isotype control.
    Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 antigen retrieval buffer EDTA pH 9.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 minutes and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.

    This image was generated using the ascites version of the product.

  • Immunocytochemistry - Anti-Mitotic proteins antibody [MPM-2] (ab14581)
    Immunocytochemistry - Anti-Mitotic proteins antibody [MPM-2] (ab14581)This image is courtesy of an anonymous Abreview

    ab14581 staining Mitotic proteins - Mitosis Marker in Human Saos-2 cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.25% Triton in PBS and blocked with 1% BSA for 1 hour at room temperature. Samples were incubated with primary antibody (1/500) for 1 hour. An Alexa Fluor® 594-conjugated Goat anti-mouse IgG polyclonal (1/250) was used as the secondary antibody.

    This image was generated using the ascites version of the product.

    See Abreview

  • Flow Cytometry (Intracellular) - Anti-Mitotic proteins antibody [MPM-2] (ab14581)
    Flow Cytometry (Intracellular) - Anti-Mitotic proteins antibody [MPM-2] (ab14581)

    Flow Cytometry analysis using anti-phospho-Ser/Thr-Pro, MPM-2 (ab14581). HeLa cells were treated with colcemid (green), or untreated (grey).

  • Western blot - Anti-Mitotic proteins antibody [MPM-2] (ab14581)
    Western blot - Anti-Mitotic proteins antibody [MPM-2] (ab14581)
    All lanes : Anti-Mitotic proteins antibody [MPM-2] (ab14581) at 2 µg

    Lane 1 : Non treated HeLa cells
    Lane 2 : Colcemid-treated HeLa cells

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 92 kDa

  • Western blot - Anti-Mitotic proteins antibody [MPM-2] (ab14581)
    Western blot - Anti-Mitotic proteins antibody [MPM-2] (ab14581)
    All lanes : Anti-Mitotic proteins antibody [MPM-2] (ab14581) at 2 µg

    Lane 1 : Non-treated HeLa
    Lane 2 : Colcemid-treated HeLa

    Predicted band size: 92 kDa

  • Immunocytochemistry - Anti-Mitotic proteins antibody [MPM-2] (ab14581)
    Immunocytochemistry - Anti-Mitotic proteins antibody [MPM-2] (ab14581)

    ab14581 (green) labeling phospho-proteins in metaphase cells. The spindle, centrosomes and centromeric regions are stained. DNA is counterstained with propidium iodide (red).

参考文献 (11)

ab14581 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。

ab14581 は 11 報の論文で使用されています。

  • Liu HY  et al. Acetylation of MORC2 by NAT10 regulates cell-cycle checkpoint control and resistance to DNA-damaging chemotherapy and radiotherapy in breast cancer. Nucleic Acids Res 48:3638-3656 (2020). PubMed: 32112098
  • Rodenfels J  et al. Heat Oscillations Driven by the Embryonic Cell Cycle Reveal the Energetic Costs of Signaling. Dev Cell 48:646-658.e6 (2019). PubMed: 30713074
  • Ruppert JG  et al. HP1a targets the chromosomal passenger complex for activation at heterochromatin before mitotic entry. EMBO J 37:N/A (2018). Flow Cyt . PubMed: 29467217
  • Machicoane M  et al. SLK-dependent activation of ERMs controls LGN-NuMA localization and spindle orientation. J Cell Biol 205:791-9 (2014). PubMed: 24958772
  • Arai A  et al. RECQL1 and WRN Proteins Are Potential Therapeutic Targets in Head and Neck Squamous Cell Carcinoma. Cancer Res 71:4598-607 (2011). PubMed: 21571861
View all Publications for this product

レビューと Q&A

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1-6 of 6 Abreviews or Q&A

Immunocytochemistry/ Immunofluorescence abreview for Anti-Mitotic proteins antibody [0.T.181] - Mitosis Marker

 Excellent
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (saos-2 human osteosarcoma)
Specification
saos-2 human osteosarcoma
Fixative
Paraformaldehyde
Permeabilization
Yes - pbs triton 0.25%
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: rt°C
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Abcam user community

Verified customer

投稿 Apr 19 2012

Question

We have check with the customer who is interested for a replacement vial for
Anti-RPLP2 antibody (ab103103) in place of ab14581 which has not worked
properly in their experiment.

Read More
Answer

Thank you for your email.

I am sorry this product had a mistake on the datasheet (which has been corrected in the meantime) and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number xxx with ab103103.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

Read More

Question

Apologies for the delay. Could we reorder the following antibody
to FOXM1 (AbCam ab83097) Rabbit polyclonal to FOXM1. Also, we would
like to take up your offer of buying three additional antibodies free
of charge. Will you send us a credit note or can we just order them
through you ?

Read More
Answer

Thank you for your email.

Yes, I can definitely send you ab83097 free of charge. For ab83097, would you like to receive 1 vial or more?

Since you had placed 3 orders for ab14581 (which we listed under the incorrect target), I can send youalso 2 more antibodies free of charge, plus the 3 I had offered you to compensate you for based on the special and unfortunate circumstances you were in.

Thus, you can either send me the catalog numbers of the antibodies that you would like to receive free of charge and I will arrange the shipment, or I can send you a credit (for the remainder ofthe 6antibodies).

Please just let me know andI will arrange for everythingaltogether.

I look forward to hear back from you.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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Question

Thanks for getting back to me. I'll send you a list
of the Abcam antibodies we have used in this study - there are quite
a lot. We will check out the alternative FOXM1 antibodies from your website and pick one or two that look suitable. Thanks for offering
three additional antibodies at no cost - our consumable funds are extremely low at the moment - that's why publishing these studies is absolutely crucial in an attempt to secure additional funding. I'll forward this e-mail to my colleague who has performed most, if not all of this work. They will then liase with you.

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Answer

Thank you for your reply.
Yes, please send me the list of your Abcam antibodies you have been using and I will check those.

I look forward to hear from your colleague to resolve this issue for you.

Thank you for your understanding.

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Question

The news of this mistake could not have come at
a worse time - we are absolutely gutted. We have spent the last
18 months performing immunohistochemistry on extremely rare tumours
(that are only available from another country) and are almost ready to submit
our manuscript. I take it from your e-mail that all of our staining
for FOXM1 using Abcam 14581 is useless ?? As I undertsand from my colleague,
who has carried out all of the IHC, we have bought several vials of
this antibody from your company, not to mention a number of other
antibody reagents (at least 6 different antibodies). I take it that
these are all OK ?? Can you please let us know which other FOXM1
antibodies are available ? My colleagu will contact you with our original
order details.

Read More
Answer

Thank you for your email.

I am extremely sorry for this oversight on our part and for the frustration and disappointment it is causing you and other customers. I completely understand your situation and sincerely apologize for this.

The antibody ab14581 is detecting mitotic proteins. The following information was sent to us from the originator of the antibody: "This antibody recognizes a variety of proteins that are phosphorylated during mitosis. This antibody does not recognize a single antigen target, but rather it reacts with phosphorylated serines or threonines when followed by a proline."
Therefore, the antibody might detect FOXM1 but it is not specific to this protein only, and staining can be seen in the cytoplasm and near centrosomes, kinetochores, and midbodies of the microtubule organizing centers. FOXM1 would be exclusively nuclear.

I am not certain which other antibodies you are using, but they are not affected by this mistake. If you please could let me know the catalog numbers, I can check if we have any additional information on these products.

I have found 3 orders from your institution in our system. I'd be happy to either replace, credit or refund these purchases. In appreciation of your circumstances, I can further offer you up to 3 additional antibodies free of charge - tohelp compensate for your losses, although I know that the time,effort and precious samples that were spent by your lab would be difficult tocompensate for, and I am so sorry about that.

As for FOXM1 antibodies: We have 6 antibodies in our catalog: ab55006, ab83097, ab100806, ab14758, ab58675, and ab14582 (phospho FOXM1, FITC-conjugated).
Please let me know which application you are specifically using (e.g. IHC fresh frozen, IHC paraffin embedded) and if the species is exclusively human. I'd be happy to assist you in finding the most suitable antibody/antibodies.

Again, please accept my sincere apologies for all this trouble.
But I look forward to hear back from you and assist you as best as I can to resolve this problem.

Read More

Question

Could you please confirm if only lot 427148 of product ab14581(MPM2, MOUSE MONOCLONAL [0.T.181] TO MPM2) is affected or any other lot is also involved?


At Abcam we aim to let our customers know everything that we do about the products in our catalogue. All the data we possess are shown on our datasheets and these are updated daily with any new information we’ve received, whether this is from customers, suppliers or our own labs. When this new information is something that may affect the quality of a product, we also contact relevant customers as a priority, to share this information and offer our assistance where necessary.

We have recently received information regarding ab14581 which you have previously purchased from us. They have informed us that there has been an error, and this antibody detects Mitotic proteins, and not FOXM1. We realize this information will impact your assessment of the data you have from this antibody, and we would like to sincerely apologize for this inconvenience.

The quality of our antibodies is very important to us, as are the feedback and data we receive. It is regrettable that the information we had on this particular antibody was not of the high standard both we and our customers expect from our products. Should you wish to accept compensation, please contact me with your original order number, and I will be pleased to arrange an immediate refund or credit note.

Should you have any further questions regarding this issue, please do contact me.

Read More
Answer

Thank you for contacting us.

The information sent is not restricted to lot427148 only, it refers to all lots.I sincerely apologize about the mistake made - which has been corrected in the meantime.

Please let me know how I can help you further, andif you need any otherinformation.

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