Overview

  • Product name
    Anti-PPM1A antibody [p6c7]
    See all PPM1A primary antibodies
  • Description
    Mouse monoclonal [p6c7] to PPM1A
  • Host species
    Mouse
  • Tested applications
    Suitable for: IHC-P, ELISA, WBmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Recombinant full length protein (Human).

  • General notes

    This product was changed from ascites to tissue culture supernatant on 28/02/19. 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.

Properties

Applications

Our Abpromise guarantee covers the use of ab14824 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 1/100.
ELISA Use at an assay dependent concentration.
WB 1/250 - 1/1000. Predicted molecular weight: 42 kDa.

Target

  • Function
    Enzyme with a broad specificity. Negatively regulates TGF-beta signaling through dephosphorylating SMAD2 and SMAD3, resulting in their dissociation from SMAD4, nuclear export of the SMADs and termination of the TGF-beta-mediated signaling.
  • Sequence similarities
    Belongs to the PP2C family.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • Protein phosphatase 2C isoform alpha antibody
    • EC 3.1.3.16 antibody
    • FLJ42306 antibody
    • IA antibody
    • MGC9201 antibody
    • Mpp alpha antibody
    • PP2C alpha antibody
    • PP2C-alpha antibody
    • PP2CA antibody
    • PP2Calpha antibody
    • PPM 1A antibody
    • PPM1A antibody
    • PPM1A_HUMAN antibody
    • PPPM1A antibody
    • Protein phosphatase 1A (formerly 2C) magnesium dependent alpha isoform antibody
    • Protein phosphatase 1A antibody
    • Protein phosphatase 1A magnesium dependent alpha antibody
    • Protein phosphatase 2C alpha antibody
    • Protein phosphatase 2C alpha isoform antibody
    • Protein phosphatase 2C isoform alpha antibody
    • Protein phosphatase IA antibody
    • Protein phosphatase, Mg2+/Mn2+ dependent, 1A antibody
    see all

Images

  • Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: PPM1A knockout HAP1 cell lysate (20 µg)
    Lane 3: HeLa cell lysate (20 µg)
    Lane 4: Jurkat cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab14824 observed at 42 kDa. Red - loading control, ab18251, observed at 52 kDa.

    ab14824 was shown to specifically react with PPM1A when PPM1A knockout samples were used. Wild-type and PPM1A knockout samples were subjected to SDS-PAGE. ab14824 diluted to 1/250 and ab18251 (loading control to alpha Tubulin) diluted to 1/10000 were incubated overnight at 4°C. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD)  preadsorbed (ab216777) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

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

  • Immunocytochemistry/ Immunofluorescence analysis of PP2C alpha/PPM1A in HeLa cells. The cell was stained with ab14824 (1:100). The secondary antibody (green) was used Alexa Fluor 488. DAPI was stained the cell nucleus (blue).

  • All lanes : Anti-PPM1A antibody [p6c7] (ab14824) at 1/1000 dilution

    Lane 1 : Jurkat cell lysate
    Lane 2 : HeLa cell lysate
    Lane 3 : K-562 cell lysate
    Lane 4 : MCF7 cell lysate
    Lane 5 : A549 cell lysate
    Lane 6 : Raji cell lysate
    Lane 7 : Mouse kidney tissue lysate
    Lane 8 : Mouse brain tissue lysate
    Lane 9 : Mouse liver tissue lysate

    Lysates/proteins at 40 µg per lane.

    Secondary
    All lanes : goat anti-mouse secondary antibody conjugated to HRP

    Predicted band size: 42 kDa

  • Western blot analysis of mouse liver cytosol extract using ab14824 at a dilution of 1/250.  Proteins were visualised using a goat anti-mouse secondary antibody conjugated to HRP and a DAB detection system. Western blot analysis of mouse liver cytosol extract using ab14824 at a dilution of 1/250. Proteins were visualised using a goat anti-mouse secondary antibody conjugated to HRP and a DAB detection system.

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

  • Anti-PPM1A antibody [p6c7] (ab14824) at 1/1000 dilution + HeLa whole cell lysate

    Secondary
    HRP conjugated anti-mouse antibody

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 42 kDa
    Observed band size: 45 kDa
    why is the actual band size different from the predicted?


    Exposure time: 10 seconds


    This image is courtesy of an Abreview submitted by Xia Lin on 2 March 2006.

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

    See Abreview

  • Ab14824 staining human colon. Staining is localised to cytoplasm.
    Left panel: with primary antibody at 4ug/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.

References

This product has been referenced in:
See all 12 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products. I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement with the order number xxx. 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 contacting Abcam regarding ab14824. I am sorry that you are experiencing difficulties with the most recent lot that you received. As this is a monoclonal antibody, I would not expect much lot to lot variation, but I would advise two modifications. I would first recommend increasing the concentration of primary antibody. We do recommend a range of 1:250 - 1:1000. Additionally, you did not indicate the amount of protein loaded, but is it possible to increase the amount loaded? Have you tested a whole cell lysate to confirm that there are no issues with the extraction kit you are using? If these suggestions do not improve your results, please do not hesitate to contact me to discuss a replacement or refund for this product per our Abpromise guarantee. Best of luck with your experiments!

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Answer

Thank you for your enquiry. I am sorry we regrettably haven’t tested the cross reactivity of ab14824 PPM1A antibody [p6c7] with other PP2C isoforms. By looking at the Western Blot image on the datasheet, we believe there’s a possibility that it is PP2C alpha specific. However, the homology between PP2C alpha and PP2C beta is 76.6% so it may cross react with the beta isoform. (the homology with PPM1N was 40%, and with other isoforms, it was around 20%) I’m sorry we couldn’t provide more information on this occasion, but if you need anything else, please contact me without hesitation.

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Application
Western blot
Sample
Human Cell lysate - other (aortic smooth muscle cells (cytoplasmic))
Loading amount
10 µg
Specification
aortic smooth muscle cells (cytoplasmic)
Gel Running Conditions
Reduced Denaturing (Bis-Tris 4-12% MOPS running buffer)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Apr 09 2008

Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa)
Specification
HeLa
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%

Xia Lin

Verified customer

Submitted Mar 02 2006

Question
Answer

Thank you for your enquiry. Unfortunately this antibody has not been tested for cross-reactivity with other isoforms. If you have any additional questions, please contact us again.

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