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Frataxin Protein Quantity Dipstick Assay Kit (ab109881)

  • Datasheet
  • Protocol Booklet
Submit a review Q&A (19)References (19)

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Sandwich ELISA - Frataxin Protein Quantity Dipstick Assay Kit (ab109881)
  • Sandwich ELISA - Frataxin Protein Quantity Dipstick Assay Kit (ab109881)
  • Sandwich ELISA - Frataxin Protein Quantity Dipstick Assay Kit (ab109881)

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Overview

  • Product name

    Frataxin Protein Quantity Dipstick Assay Kit
  • Sample type

    Whole Blood
  • Assay type

    Sandwich (quantitative)
  • Species reactivity

    Reacts with: Human
  • Product overview

    ab109881 is used to rapidly quantify frataxin protein levels from human sample materials. Purification of mitochondria is not necessary for the performance of this assay. Based on the immunologic sandwich assay, the kit utilizes two monoclonal antibodies (mAbs) specific to different antigens present on the mature form of frataxin. One antibody is immobilized on the nitrocellulose membrane of the dipstick in a thin line perpendicular to the length of the dipstick while the other is gold-conjugated which gives a visual signal. When frataxin is present in the sample, a red-colored line appears on the dipstick at the site of the anti-frataxin mAb immobilized on the membrane. The signal intensity is directly related to the level of frataxin in the sample. The signal intensity is best measured by a dipstick reader or may be analyzed by another imaging system.

  • Notes

    All components are stable in their provided containers at room temperature out of direct sunlight.

    After diluting the 10X Blocking Buffer to 2X, store at 4°C.

    For long-term storage, all buffers can be stored at 4°C.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Reagents

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 30 tests
    Buffer B (10X Blocking solution) 1 x 0.4ml
    Dipsticks 1 x 30 units
    Extraction Buffer (ab260490) 1 x 15ml
    Gold-conjugated antibody (dried in microplate wells) 1 x 30 tests
    Wash buffer 1 x 2ml
  • Research areas

    • Signal Transduction
    • Metabolism
    • Mitochondrial
    • Neuroscience
    • Neurology process
    • Neurodegenerative disease
    • Other
    • Signal Transduction
    • Metabolism
    • Vitamins / Minerals
    • Metabolism
    • Pathways and Processes
    • Mitochondrial Metabolism
    • Mitochondrial markers
  • Alternative names

    • FRDA
  • Database links

    • Entrez Gene: 2395 Human

    Applications

    The Abpromise guarantee

    Our Abpromise guarantee covers the use of ab109881 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
    Sandwich ELISA
    Use at an assay dependent dilution.
    Notes
    Sandwich ELISA
    Use at an assay dependent dilution.

    Images

    • Sandwich ELISA - Frataxin Protein Quantity Dipstick Assay Kit (ab109881)
      Sandwich ELISA - Frataxin Protein Quantity Dipstick Assay Kit (ab109881)
      An example using ab109881 to quantify frataxin levels from ß -lymphocyte cell culture samples derived from control individuals and Friedreich's Ataxia (FA) patients. Based on the above standard curve values, of protein extract for the controls and FA patient samples. (Note: for a statistical analysis, it is preferred to use two dipsticks for each sample; intra-assay CV's are typically <=10%.) For this analysis, the FA patients had between 550 - 925 GAA repeats on the smaller allele. Using GraphPad software, the signal intensity from the standard curve was interpolated and the relative amount of frataxin in the patients, as compared to the controls, was determined. Based on the above analysis, the patient samples had between 13% and 20% of frataxin levels compared to the control.
    • Sandwich ELISA - Frataxin Protein Quantity Dipstick Assay Kit (ab109881)
      Sandwich ELISA - Frataxin Protein Quantity Dipstick Assay Kit (ab109881)
      An example using ab109881 to quantify frataxin levels from ß -lymphocyte cell culture samples derived from control individuals and Friedreich's Ataxia (FA) patients. Shown is a 1:2 dilution series using a positive control sample. Approximately 6 to 8 dipsticks are suitable for covering the entire working range and the blank for background levels. In this example the dilution series starts with 4 µg of control sample. A one-site hyperbola line was generated for best-fit analysis using GraphPad.
    • Sandwich ELISA - Frataxin Protein Quantity Dipstick Assay Kit (ab109881)
      Sandwich ELISA - Frataxin Protein Quantity Dipstick Assay Kit (ab109881)
      Dipstick assays use the well-established lateral flow concept, whereby capture antibodies are striped onto nitrocellulose membrane and a Whatman paper wicking pad draws the sample through the antibody bands. Detector antibodies, conjugated to gold, are dried in the wells of a 96-well plate. Sample is added to the well, the dipstick inserted, and within minutes the line for each target is revealed as the protein-detector antibody-gold complex binds with the capture antibodies. Multiplexing dipstick assays have multiple target protein lines. A positive control goat anti-mouse antibody line is included on all assays to ensure that adequate wicking of the sample occurred.

    Protocols

    • Protocol Booklet

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet download

      Download

    References (19)

    Publishing research using ab109881? Please let us know so that we can cite the reference in this datasheet.

    ab109881 has been referenced in 19 publications.

    • Mikaeili H  et al. FAST-1 antisense RNA epigenetically alters FXN expression. Sci Rep 8:17217 (2018). PubMed: 30464193
    • Cherif K  et al. Increased Frataxin Expression Induced in Friedreich Ataxia Cells by Platinum TALE-VP64s or Platinum TALE-SunTag. Mol Ther Nucleic Acids 12:19-32 (2018). PubMed: 30195758
    • Crombie DE  et al. Friedreich's ataxia induced pluripotent stem cell-derived cardiomyocytes display electrophysiological abnormalities and calcium handling deficiency. Aging (Albany NY) 9:1440-1452 (2017). PubMed: 28562313
    • Chapdelaine P  et al. Development of an AAV9 coding for a 3XFLAG-TALEfrat#8-VP64 able to increase in vivo the human frataxin in YG8R mice. Gene Ther 23:606-14 (2016). Human . PubMed: 27082765
    • Wedding IM  et al. Friedreich ataxia in Norway - an epidemiological, molecular and clinical study. Orphanet J Rare Dis 10:108 (2015). Human . PubMed: 26338206
    View all Publications for this product

    Customer reviews and Q&As

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

    1-10 of 19 Abreviews or Q&A

    Question

    hello,

    here is a picture of what I mean.

    1-2-3-4 are samples to test

    5-6-7 are negative controls

    8-9 are positive controls

    they are all mouse muscle extracts. I put 40ng total proteins in the well.

    thank you to give me your opinion about the GAM control!

    Read More

    Abcam community

    Verified customer

    Asked on Apr 03 2013

    Answer

    Thank you for this information.

    The control lines are not all the same between kits. Within a single kit the control can also change –it’s possible if the signal from the sample is very strong this can deplete the mount gold labeled detector antibody and would weaken the positive control band.The positive control band is qualitative and should not be measured or assessed quantitatively.

    Please let me know if you have any further questions.

    Read More

    Abcam Scientific Support

    Answered on Apr 03 2013

    Question

    Thank you for the information,

    I use dipsticks on mice tissue extracts the are treated or not with human frataxin. I detect a strong band in my hFXN positive tissues, but I also detect a band in my untreated tissues.

    Are-you sure dipstick is human specific? It seems to me that it cross-react with mouse frataxin...

    Read More

    Abcam community

    Verified customer

    Asked on Mar 19 2013

    Answer

    Thank you for contacting us.


    We have done pretty extensive testing with the antibodies used in MSF31 to show that is human specific - the detector antibody is human only.

    So If there is a band in mouse I would be concerned about background binding, I would recommend reducing the amount of sample for both treated and untreated.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More

    Abcam Scientific Support

    Answered on Mar 19 2013

    Question

    Hello,

    are antibodies included in the dipstick the same as the one in the elisa kit?


    Read More

    Abcam community

    Verified customer

    Asked on Mar 18 2013

    Answer

    Thank you for contacting us.

    No the antibodies in these two kits are not the same- dipsticks ab109881 contains antibodies that are human specific these are the same human specific antibodies can be found in ELISA kit ab110173.

    However for the kit mentioned ab115346 we developed new antibodies that also cross react with multiple other species. At this time we have not made the multispecies Frataxin dipstick.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More

    Abcam Scientific Support

    Answered on Mar 18 2013

    Question

    Phone call requesting confirmation that the ab109881 is compatible with ab109883

    Read More

    Abcam community

    Verified customer

    Asked on Jul 31 2012

    Answer

    Thank you for contacting us yesterday in regards to the Frataxin and Pyruvate dehydrogenase Quantity Dipstick kits (ab109881 and ab109883 respectively).

    As discussed over the phone, I contacted MitoSciences to discuss if the kits would be compatible with the experiment you are hoping to perform and if this would be a valuable way of normalising the Frataxin values collected.

    I have the following information to share:

    The buffer A from ab109881 kitis the same component as thebuffer A from the ab109883 kit. The samples prepared by eluting with this buffer should therefore be compatible for measurement using either kit.

    In regards to the questions whether the Pyruvate dehydrogenase kit is likely to serve as a good normalising standard of your samples this is a less straight forward answer. The PHD Protein Quantity Kit (ab109883) will allow you to measure the PHD quantity present in yoursampleswith sufficient accuracy to use as a normalising value. However,even though Pyruvate dehydrogenase is ubiquitous andtheproteinisencoded by “housekeeping genes”, deciding on the perfect control will vary a lot on the population you are looking at.

    It would be worthwhile to testwhether or not PDH is a good control protein for your particular sample. One way of doing this is by doing a longitudinal study on a small control cohort to determine: (1) Does PDH change significantly from subject to subject and (2) does PDH change significantly through time in the same cohort.If the answer to both is NO, then it could be used as a control protein.





    I hope this has sufficiently answered your questions but please do not hesitate to contact us again if you have any further questions.

    Read More

    Abcam Scientific Support

    Answered on Jul 31 2012

    Question

    Dear Abcam / Mitosciences representative

    We have been using the Frataxin dipstick assay (ab109881 -MSF31) for a long time now and now actively using the kit in a clinical trial.

    We now want to use an additional dipstick in order to be able to normalise the samples. Being a housekeeping gene, we thought that pyruvate dehydrogenase would be a good candidate. Since mitosciences already has dipsticks for this protein, we would like to test if this will work. Here, the key question is, are the buffers in the FXN dipstick assay compatible with the PDH dipsticks? I am asking this because we elute the total proteins (extracted with trizol first) in FXN dipstick buffer A and we want to use the same sample to test for the PDH as well.

    Could you please inform me about this? Also, is it possible for you to send us some samples for the PDH dipsticks (ab109883-MSP31) for us to test? Being a regular customer, I hope you will consider this.

    Kind regards

    Read More

    Abcam community

    Verified customer

    Asked on Jul 25 2012

    Answer

    Thank you for your enquiry.

    For the sandwich ELISA dipstick ab109881, the buffers are the same. However for the PDH activity dipstick ab109883 they are very different.

    I am sorry that as we have over 80 0000 products it is not possible to send test samples. I would like to reassure you that we can provide our 6 months guarantee for products used in the tested applications and species specified on the individual datasheets. If they do not work in the tested applications and species, we can provide a refund, credit note or free of charge replacement.

    I hope this will be helpful. If you have any further questions, please do not hesitate to contact us.

    Read More

    Abcam Scientific Support

    Answered on Jul 25 2012

    Question

    Dear Mitosciences/Abcam representative,

    I am working as a post doc in a major research lab that is focused on Friedreich's Ataxia. At the moment, we are performing a clinical trial during which we are using mitosciences Frataxin dipstick assay. Having used this commercial assay for a long time now, I need to ask you the following questions:

    1) I am using ˜5 million peripheral mononuclear blood cells per sample. I do Trizol extraction first in order to get RNA and protein pellet from the same sample. After I obtain the protein pellet, I try to dissolve it in 500 microlitre of mitosciences Dipstick buffer A. However, the pellet never completely dissolves. I tried much smaller cell numbers and higher volumes of the buffer A but still there is always an insoluble material. Having said this though, I could detect that I always have a protein concentration (by BCA assay) and the assay seems to work fine to detect various FXN ranges among healthy and affected individuals. Can you please comment about this? Is this normal? Can we improve the solubility of the pellet?

    2) Also, we want to confirm the dipstick results with Mass Spectrometry as well. However, we are concerned that the Buffer A may have incompatible ingredients. I understand that it may be your policy not to disclose the full formula of the dipstick buffer A but could you please let us know about the composition of this buffer?

    Your kind considerations will be deeply appreciated.

    Kind regards

    Read More

    Abcam community

    Verified customer

    Asked on Jul 03 2012

    Answer

    Thank you for contacting us.

    Please find below the answers from my colleagues from Abcam-MitoSciences regarding the kit ab109881:

    1)This is normal. The detergent provided with the kit is a very mild detergent and which does not solubilize much of the cell nucleus, however all other organelle’s protein will solublize. I think the assay is working well and there is no need to improve the solubility.

    2) Here are the components of buffer A = Lauryl maltoside, Hepes, NaCl

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More

    Abcam Scientific Support

    Answered on Jul 03 2012

    Question

    I am getting around 450mAbs by using 1ng of rFxn, and around 250-290mabs by using 0.5ng rFxn,

    let me know what do you suggest to increase the absorbance, will call you tomorrow.

    Read More

    Abcam community

    Verified customer

    Asked on Apr 03 2012

    Answer

    Thank you for contacting us.

    I assume the samples you are using are just recombinant human Frataxin, not a cell lysate?

    To increase the signal it is suggested to add more sample to another dipstick.

    What type of dipstick reader are you using?
    How old is the kit you are using?

    I look forward to hear back from you and assist you further.

    Read More

    Abcam Scientific Support

    Answered on Apr 03 2012

    Question

    What is the absorbance you would get for the recombinant proteinusing these kits?

    Read More

    Abcam community

    Verified customer

    Asked on Mar 30 2012

    Answer

    Thank you for contacting us.

    This is the information, we received from the lab:

    The exact number for ab109881 will vary from batch to batch of kit and there will be a small variation depending on how old the kit is. However, roughly speaking, the customer should expect the following milliabsorbance from the dipstick reader =

    1ng/dipstick = 600 milliabsorbance
    0.2ng/dipstick = 200 milliabsorbance
    25pg/dipstick = 40 milliabsorbance


    For dipsticks we have a dedicated reader: ab117066 (https://www.abcam.com/Dipstick-Reader-ab117066.html).


    As for ab110173, the protocol can be found here: https://www.abcam.com/ps/products/110/ab110173/documents/ab110173%20Frataxin%20Protein%20Quantity%20ELISA%20Kit%20(Website).pdf

    or on the online datasheet for the product. The OD readings are to be performed at 450 nm.

    I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Read More

    Abcam Scientific Support

    Answered on Mar 30 2012

    Question

    Can you send the link or PDF for a Frataxin dipstick and plate assays? What are the detection limits, sample sizes and protocol, etc.

    Thanks,

    Read More

    Abcam community

    Verified customer

    Asked on Mar 23 2012

    Answer

    Thank you for your enquiry.

    I am pleased to help. I assume you are referring to the following frataxin kits from our catalog:

    ab109881 Frataxin Protein Quantity Dipstick Assay Kit
    Reacts with: Human
    https://www.abcam.com/Frataxin-Protein-Quantity-Dipstick-Assay-Kit-ab109881.html (or use the following: https://www.abcam.com/Frataxin-Protein-Quantity-Dipstick-Assay-Kit-ab109881.html).

    ab115346 Frataxin ELISA Kit
    Tested in:ELISA
    Reacts with: Human, Mouse, Rat, Cow, Dog
    https://www.abcam.com/FANCI-antibody-ab15346.html (or use the following: https://www.abcam.com/FANCI-antibody-ab15346.html).

    ab110173 Frataxin Protein Quantity Microplate Assay Kit
    Tested in: sELISA
    Reacts with: Human
    https://www.abcam.com/Rat-Frataxin-Profiling-ELISA-Kit-ab110173.html (or use the following: https://www.abcam.com/Rat-Frataxin-Profiling-ELISA-Kit-ab110173.html).

    The protocols for these can be found on the online datasheets. Please click on the datasheet links above for more details. On the datasheet, there is a 'protocols' tab. Click on this, and then click on the Protocols booklet link under 'specific protocols'. There is also a link to the protocol on the main datasheet page.

    The limits of detection for all kits is very similar, around 0.1 ng/mL for frataxin protein.

    For samples working range we suggest the following:
    Typical Working Ranges:

    Tissue extract (e.g. Liver) 10 - 250 μg/mL
    Whole cultured cell extract 20 - 500 μg/mL
    Pure Frataxin protein 0.1 -12 ng/mL

    I hope this information will be helpful to you. If you have any further questions, please do not hesitate to contact me.

    Read More

    Abcam Scientific Support

    Answered on Mar 23 2012

    Question

    Missing buffers B and C from this kit.

    Read More

    Abcam community

    Verified customer

    Asked on Mar 22 2012

    Answer

    Thank you for your calls this week and for your patience.

    I attempted to return your call today but the number that I have in my notes is still not correct. Could you send me the number where I can reach you?

    I'm still in the process of seeing whether we can sell these kit components, however it is possible that we will not be able to do so. I have asked the lab scientists if there are any other buffers that could be substituted, as buffers B and C are the wash and blocking buffers so there might be other compatible commercial buffers.

    I will be out of the office tomorrow, but I will follow-up with you on Monday and I hope to have this resolved by then. Please let me know if you have any questions or if there is anything else that we can do for you.

    Read More

    Abcam Scientific Support

    Answered on Mar 22 2012

    1-10 of 19 Abreviews or Q&A

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