Overview

  • Product name
    Anti-Mitotic proteins antibody [MPM-2]
  • Description
    Mouse monoclonal [MPM-2] to Mitotic proteins
  • Host species
    Mouse
  • Specificity
    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.
  • Tested applications
    Suitable for: IHC-P, ICC/IF, IP, WB, ICC, ELISAmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: a wide range of other species
  • Immunogen

    Tissue/ cell preparation (Mitotic human HeLa cell cytosolic lysate).

  • Positive control
    • Colcemid treated HeLa cell lysate.
  • General notes

    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.

Properties

Applications

Our Abpromise guarantee covers the use of ab14581 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
IHC-P Use a concentration of 2 µg/ml.
ICC/IF Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
WB Use a concentration of 0.5 - 2 µg/ml. Predicted molecular weight: 92 kDa.
ICC Use a concentration of 5 - 15 µg/ml.
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).

Target

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

Images

  • 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.
  • 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.

    See Abreview

References

This product has been referenced in:
  • 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 . Read more (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). Read more (PubMed: 24958772) »
See all 8 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

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.

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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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Abcam guarantees this product to work in the species/application used in this Abreview.
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

Abcam user community

Verified customer

Submitted Apr 19 2012

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.

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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.

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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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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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