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    products/assay-kits/propidium-iodide-ab14083.pdf

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Cell Biology Apoptosis Annexin V Reagents
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Propidium Iodide (ab14083)

  • Datasheet
Submit a review Q&A (9)References (34)

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Abpromise

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Flow cytometry: ab14083
  • Immunofluorescence analysis using ab14083

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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 (λ= 536/617). Cells will only be stained if the membrane has been permeated, either naturally (non-viable cells) or with detergents (for fluorescent staining).


    For Flow Cytometry analysis: PI staining can be monitored in FL2 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.

  • Notes

    This product is manufactured by BioVision, an Abcam company and was previously called 1056 Propidium Iodide. 1056-1 is the same size as the 1 ml size of ab14083.

  • Tested applications

    Suitable for: FM, Flow Cytmore details

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components Identifier 1 ml
    Propidium Iodide (1mg/ml) ab14083 1 x 1ml
  • Research areas

    • Cell Biology
    • Apoptosis
    • Annexin V Reagents
    • Kits/ Lysates/ Other
    • Tools and Reagents
    • IHC Tools/ Reagents
    • Kits/ Lysates/ Other
    • Kits
    • Apoptosis Kits
    • Apoptosis Assay Kits
    • Cell viability
    • Kits/ Lysates/ Other
    • Kits
    • Cell Damage Kits
    • DNA damage and cell cycle arrest
    • Kits/ Lysates/ Other
    • Kits
    • Cell Damage Kits
    • Cell Damage
    • Cancer
    • Cell Death
    • Apoptosis
    • Apoptosis Markers
    • Annexin V
    • Annexin V Reagents
  • Relevance

    Propidium Iodide (PI) (MW=668.4 Da) is an intercalating agent and a fluorescent molecule which is membrane impermeant and generally excluded from viable cells. Upon entering cells, PI will bind to DNA and RNA by intercalating between bases. Once bound to the nucleic acids, its fluorescence is enhanced 20- to 30-fold. Exmax= 536nm / Emmax= 617 nm.

Associated products

  • Related Products

    • FCCP, mitochondrial oxidative phosphorylation uncoupler (ab120081)
    • Cycloheximide, Protein synthesis inhibitor (ab120093)
    • SB431542, ALK inhibitor (ab120163)
    • Z-VAD(OH)-FMK, Irreversible general caspase inhibitor (ab120382)
    • Z-VAD(OMe)-FMK, Cell permeable, irreversible pan-caspase inhibitor (ab120487)
    • Dorsomorphin (Compound C), AMP-kinase inhibitor (ab120843)
    • Annexin V/ANXA5-FITC Apoptosis Detection Reagent (500X) (ab14082)
    • 10X Annexin V/ANXA5 Binding Buffer (ab14084)
    • Annexin V-FITC Apoptosis Staining / Detection Kit (ab14085)
    • Annexin V-Cy3 Apoptosis Staining / Detection Kit (ab14142)
    • Calcein AM, fluorescent dye for cell viability (ab141420)
    • Annexin V/ANXA5-Cy3 Apoptosis Staining / Detection Reagent (ab14143)
    • Annexin V-Cy3 Apoptosis Staining / Detection Kit with SYTOX (ab14144)
    • Annexin V/ANXA5-Cy5 Apoptosis Staining / Detection Reagent (ab14147)
    • Annexin V-Cy5 Apoptosis Staining / Detection Kit (ab14150)
    • Annexin V-EGFP Apoptosis Staining / Detection Reagent (ab14152)
    • Annexin V-EGFP Apoptosis Staining / Detection Kit (ab14153)
    • Annexin V/ANXA5-PE Apoptosis Staining / Detection Reagent (ab14154)
    • Annexin V-PE Apoptosis Staining / Detection Kit (ab14155)
    • Annexin V/ANXA5-Biotin Apoptosis Staining / Detection Reagent (ab14165)

Applications

The Abpromise guarantee

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.
Flow Cyt
Use at an assay dependent concentration.
Notes
FM
Use at an assay dependent concentration.
Flow Cyt
Use at an assay dependent concentration.

Images

  • Flow cytometry: ab14083
    Flow cytometry: ab14083Image from Wang et al., PLoS One., 10(9). Fig 3a & c. doi:10.1371/journal.pone.0137712 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    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).

     

  • Immunofluorescence analysis using ab14083
    Immunofluorescence analysis using ab14083Image from Melia MM et al ., Plos One.,29 (8). Fig 1b DOI: 10.1371/journal.pone.0106281. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/

    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). 

     

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

Datasheets and documents

  • Datasheet download

    Download

References (34)

Publishing research using ab14083? Please let us know so that we can cite the reference in this datasheet.

ab14083 has been referenced in 34 publications.

  • Jiang T  et al. Application of Bone Marrow Mesenchymal Stem Cells Effectively Eliminates Endotoxemia to Protect Rat from Acute Liver Failure Induced by Thioacetamide. Tissue Eng Regen Med 19:403-415 (2022). PubMed: 35122584
  • Feehan DD  et al. Natural killer cells kill extracellular Pseudomonas aeruginosa using contact-dependent release of granzymes B and H. PLoS Pathog 18:e1010325 (2022). PubMed: 35202434
  • Eichelmann AK  et al. Mutant p53 Mediates Sensitivity to Cancer Treatment Agents in Oesophageal Adenocarcinoma Associated with MicroRNA and SLC7A11 Expression. Int J Mol Sci 22:N/A (2021). PubMed: 34074015
  • Ceder S  et al. A thiol-bound drug reservoir enhances APR-246-induced mutant p53 tumor cell death. EMBO Mol Med 13:e10852 (2021). PubMed: 33314700
  • Zhang H  et al. KCNQ1OT1 regulates the retinoblastoma cell proliferation, migration and SIRT1/JNK signaling pathway by targeting miR-124/SP1 axis. Biosci Rep 41:N/A (2021). PubMed: 33345272
View all Publications for this product

Customer reviews and Q&As

Show All Reviews Q&A
Submit a review Submit a question

1-9 of 9 Abreviews or Q&A

Question

Propidium iodide flow cytometry protocol

Read More

Abcam community

Verified customer

Asked on Oct 21 2016

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.

Read More

Abcam Scientific Support

Answered on Oct 21 2016

Question

Hello  we are interested in participating the AbTrial program We will purchase two antibodies this time:  1. Sprouty-1: Ab56670 2. Sprouty-2: Ab60719  

Read More

Abcam community

Verified customer

Asked on Nov 09 2012

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.

Read More

Abcam Scientific Support

Answered on Nov 09 2012

Question

Hi ,
Many thanks for your reply, excellent news. Do you have a suggested protocol that i could have and can it be used in conjunction with Annexin V to detect apoptosis? Sorry to ask a lot of questions.
Kind Regards,

Read More

Abcam community

Verified customer

Asked on May 30 2012

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.

Read More

Abcam Scientific Support

Answered on May 30 2012

Question

rormalin fixed paraffin embedded tissues. Will this be suitable for us?
Many Thanks,

Read More

Abcam community

Verified customer

Asked on May 29 2012

Answer

rr/>

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.

Read More

Abcam Scientific Support

Answered on May 29 2012

Question

Dear Sir/Madam,

I am enquiring about your Propidium Iodide (ab14083). Is this product suitable for using on fixed tissue samples? I would like to identifiy necrosis by fluorescence microscopy in my wound tissue samples (mouse).
Kind Regards,

Read More

Abcam community

Verified customer

Asked on May 25 2012

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.

Read More

Abcam Scientific Support

Answered on May 25 2012

Question

habe den Assay ohne Ribonuclease gemacht, sehe keine individuellen peaks, nur einen. Wie wichtig ist dieser Schritt?
https://www.abcam.com/index.html?pageconfig=resource&rid=13432

Read More

Abcam community

Verified customer

Asked on Mar 23 2012

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.

Read More

Abcam Scientific Support

Answered on Mar 23 2012

Question

Abcam, I am using abcam Propidium Iodide (ab14083) staining of cells to assess DNA cell cycle according to https://www.abcam.com/index.html?pageconfig=resource&rid=13432 After singlets gating and gate out the debris, I have the histogram as below. G0/G1, S, G2/M didn’t change much but I found the population close/overlaid with y-axis changed significantly. My question is: is that population sub G1 or debris? Sample #1: time 0 hr Sample #2: time 72 hr

Read More

Abcam community

Verified customer

Asked on Dec 23 2011

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.

Read More

Abcam Scientific Support

Answered on Dec 23 2011

Question

Will this dye measure the subG0 phase?

Read More

Abcam community

Verified customer

Asked on Dec 15 2011

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.

Read More

Abcam Scientific Support

Answered on Dec 15 2011

Question

I am seeing alot of staining in the zebrafish yolk. Why?

Read More

Abcam community

Verified customer

Asked on Jun 12 2006

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.

Read More

Abcam Scientific Support

Answered on Jun 15 2006

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