Overview

  • Product name
    Propidium Iodide
  • Product overview

    Propidium Iodide (PI) binds to double-stranded DNA. PI cannot cross intact plasma membrane and therefore will only be present in DNA of cells where the plasma membrane has been compromised/ permeabilized.

    For Microscopy analysis: PI can be viewed using rhodamine(red) filter (?= 536/617). Cells will only be stained if the membrane has been permeated, either naturally (non-viable cells) or with detergents (for fluorescent staining).

    Flow Cytometry analysis: PI staining can be monitored in FL2 (DNA content) or FL3 (viability) channel.

    If used together as control for Annexin V assays ab14082, ab14083 or ab14152, PI should be diluted to 250 µg/ml solution (in PBS) prior use and added as 1 µl/ Annexin V Assay (0.25µg/assay).

    Visit our FAQs page for tips and troubleshooting.

  • Tested applications
    Suitable for: FMmore details

Properties

Applications

Our Abpromise guarantee covers the use of ab14083 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
FM Use at an assay dependent concentration.

Images

  • Vero-DogSLAM (VDS) cells and Vero (African Green Monkey kidney) cells were infected with wtCDV. Cells were fixed, permeabilised and stained with SSPE serum and rabbit anti-human FITC; nuclei were stained using ab14083 propidium iodide. Images were taken using a Nikon Eclipse TE2000-U UV microscope (x400). 

     

  • Wang et al. used ab14083 to investigate an optimized platform for maintaining proliferation of giant panda mesenchymal stem cells (MSCs). MSCs were fixed in 70% alcohol and 30% PBS at 4°C for 1 hour. Cells are then incubated in the dark with PBS, 20µg/ml propidium iodide (ab14083) and 1% RNaseA for 30 minutes at 37°C. Cell samples are resuspended in PBS and analyzed using FACS Calibur flow cytometry.

    Flow cytometry analysis shows the cells are at different cell cycle stages in different concentrations (0ng/ml and 5ng/ml respectively) of basic fibroblast growth factor (bFGF).

     

References

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

Customer reviews and Q&As

1-9 of 9 Abreviews or Q&A

Question
Answer

Here is a link to the PI staining protocol I found, https://www.abcam.com/protocols/flow-cytometric-analysis-of-cell-cycle-with-propidium-iodide-dna-staining


It suggests using a solution of 50ug/mL, so dilute the amount of the 1mg/mL stock that you anticipate needing 1/20 to get it to 50ug/mL. PBS is fine for the diluent.

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Answer

I am very pleased to hear you would like to accept our offer and test these products in flow cytometry. Each code will give you the value listed off your next order before the expiration date. To redeem this offer, please submit an Abreview for flow cytometry 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. Abcam also carries other products essential for Flow Cytometry such as isotype controls which confirm that the primary antibody binding is specific and not results of non-specific Fc receptor binding or other protein interactions. The isotype control should match the species, isotype, clonality and conjugation of the primary antibody. In this case the isotype control ab56670 is: Mouse IgG2a, kappa monoclonal [MG2a-53] - Isotype control (ab18414) and for ab60719: Mouse IgG1, kappa monoclonal [MG1-45] - Isotype control (ab18447). More information about these isotype controls may be found at the following locations: Mouse IgG2a, kappa monoclonal [MG2a-53] - Isotype control (ab18414): https://www.abcam.com/mouse-igg2a-kappa-monoclonal-mg2a-53-isotype-control-ab18414.html Mouse IgG1, kappa monoclonal [MG1-45] - Isotype control (ab18447): https://www.abcam.com/mouse-igg1-kappa-monoclonal-mg1-45-isotype-control-ab18447.html If testing live cells we also recommend cell viability controls such as 7-AAD or Propidium Iodide. Propidium Iodide (PI) binds to double-stranded DNA. PI cannot cross intact plasma membrane and therefore will only be present in DNA of cells where the plasma membrane has been compromised/ permeabilized. We do carry PI and offer a 7-AAD Cell-Mediated Cytotoxicity Assay Kit. Propidium Iodide: https://www.abcam.com/Propidium-Iodide-ab14083.html 7-AAD Kit: https://www.abcam.com/7-AADCFSE-Cytotoxicity-Assay-Kit-ab133073.html 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/abtrial.

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Answer

Thank you for your reply.

Both Annexin V-EGFP Apoptosis Detection Reagents (ab14152 and ab14082) contain a protocol booklet in their datasheets. These are the recommended protocols to use along with the Propidium Iodide reagent.

Please find below the link to these booklets:

https://www.abcam.com/ps/products/14/ab14082/documents/ab14082%20Annexin%20V%20FITC%20Apoptosis%20Detection%20Reagent%20(Website).pdf

https://www.abcam.com/ps/products/14/ab14152/documents/ab14152%20Annexin%20V%20EGFP%20Apoptosis%20Detection%20Reagent%20(Website).pdf

If you had any doubt or suggestions regarding the protocols or any other products, please do not hesitate to contact us back. We are more than happy to help.

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Question
Answer

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I confirm the product Propiduim Iodide ab14083 can definitely be used with paraffin-embedded, formalin-fixed tissues.

Thank you for your comprehension, and please do not hesitate to contact us back for further assistance.

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Answer

Thank you for contacting us.

Yes, Propidium Iodide ab14083 can definitely be used with fixed tissue samples.

Please do not hesitate to contact us if you need any more advice or information.

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Answer

Vielen Dank für Ihre Geduld.

Wie versprochen habe ich mich nach den aktuellen Details des Propidiumiodid-Staining Protokolls erkundigt – an diesem seit Jahren bewährten Protokoll hat sich nichts geändert. Ein Mitarbeiter von Derek Davies hat bestätigt, dass der Verdau mit RNase essentiell ist, um zu gewährleisten, dass nur DNA angefärbt wird. Der RNA-Gehalt von Zellen variiert immer sehr stark; unter Umstaenden kann das RNA-Signal dann so stark sein, dass das DNA-Signal überlagert wird. Für akkurate Ergebnisse ist dieser Schritt also unabdingbar.

Wenn Sie aus experimentellen oder anderen Gründen keinen RNA-Verdau durchführen möchten, müssten Sie auf einen Farbstoff zurückgreifen, der (hinreichend) DNA-spezifisch ist, wie zum Beispiel DRAQ5 (ab108410) oder DAPI. Für diese benötigen Sie allerdings aufgrund der spektralen Eigenschaften (Ex 646 nm/Em 681 nm /697 nm intercalated with dsDNA bzw. Ex 358nm/Em 461 nm) entsprechende (UV-)Laser und Filter.

PI weist bei 605 nm ein Emissionsmaximum auf, so dass es auf einem Calibur entweder in FL2 (585/42BP Filter) oder FL3 (650LP Filter) gemessen werden kann. Auf LSR-Cytometern kann PI mit dem 610/10 Filter, dem 575/20 Blue-Filter oder dem 640/20 Yellow-Filter gemessen werden. Für die meisten Säugerzellen liegen typische Spannungen für PI um 400 V (Calibur) beziehungsweise 300 V (LSR).

Wenn Sie keine individuellen Peaks für die einzelnen Zellzyklusphasen detektieren, kann dies auch ein Hinweis darauf sein, dass die Konzentration von PI zu hoch ist. In diesem Fall kann eine Titration helfen, die optimale Konzentration für Ihre experimentellen Gegebenheiten zu ermitteln.

Ich hoffe, diese Informationen helfen Ihnen weiter und wünsche Ihnen viel Erfolg für Ihr Projekt. Bitte zögern Sie nicht, sich wieder bei uns zu melden, falls Sie weitere Fragen haben.

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Answer

Thank you for sending your data, it was very helpful in this case. Since the first peak is much higher, I suspect it should be a combination of sub G1 and cell debris. Usually if it is cell debris, it should just be at the bottom and there shouldn’t be a peak. But in this case probably the presence of increased sub G1 cells or a possible contamination could yield such a result. Please let me know if there is anything else I can help you with.

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Question
Answer

 Thank you for calling Abcam earlier. I am going to email the lab about your query and I just wanted to make sure I had written it down correctly, were you asking: "Will Propidium Iodide label the sub G0 phase" Is that correct? If so, I will go ahead and forward you question to the lab. I look forward to your reply.

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Question
Answer

Thank you for your enquiry and your patience. I do not have any specific advice on why you see staining in the yolk. The best resource is probably the protocols you can find in the zebrafish book at www.zfin.org I believe that methanol washes may be a way to remove the staining in the yolk, but another zebrafish expert may have more information than me. I did not find any specific information addressing this problem in my old protocols. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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