Anti-LONP1/Lon antibody (ab103809)

Reviews (1)Q&A (5)References (15)

Overview

  • Product name

    Anti-LONP1/Lon antibody
    See all LONP1/Lon primary antibodies

  • Description

    Rabbit polyclonal to LONP1/Lon

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WBmore details

  • Species reactivity

    Reacts with: Mouse, Human

  • Immunogen

    Recombinant full length protein corresponding to Human LONP1/Lon aa 1-959.

  • Positive control

    • Human kidney, Mouse kidney, HepG2, transfected 293T cell line

  • General notes

    Protein previously labeled as LONP1.

Properties

Applications

Our Abpromise guarantee covers the use of ab103809 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
WB Use a concentration of 1 - 5 µg/ml. Predicted molecular weight: 106 kDa.

Target

  • Function

    ATP-dependent serine protease that mediates the selective degradation of misfolded, unassembled or oxidatively damaged polypeptides as well as certain short-lived regulatory proteins in the mitochondrial matrix. May also have a chaperone function in the assembly of inner membrane protein complexes. Participates in the regulation of mitochondrial gene expression and in the maintenance of the integrity of the mitochondrial genome. Binds to mitochondrial promoters and RNA in a single-stranded, site-specific, and strand-specific manner. May regulate mitochondrial DNA replication and/or gene expression using site-specific, single-stranded DNA binding to target the degradation of regulatory proteins binding to adjacent sites in mitochondrial promoters. Endogenous substrates include mitochondrial steroidogenic acute regulatory (StAR) protein.

  • Tissue specificity

    Duodenum, heart, lung and liver, but not thymus.

  • Involvement in disease

    CODAS syndrome

  • Sequence similarities

    Belongs to the peptidase S16 family.
    Contains 1 Lon domain.

  • Cellular localization

    Mitochondrion matrix.
  • Information by UniProt

  • Database links

    • Alternative names

      • hLON antibody
      • hLON ATP dependent protease antibody
      • LON antibody
      • lon peptidase 1, mitochondrial antibody
      • LON protease antibody
      • Lon protease homolog antibody
      • Lon protease like protein antibody
      • Lon protease-like protein antibody
      • LONHs antibody
      • LONM_HUMAN antibody
      • LONP antibody
      • Lonp1 antibody
      • MGC1498 antibody
      • mitochondrial antibody
      • Mitochondrial ATP dependent protease Lon antibody
      • Mitochondrial ATP-dependent protease Lon antibody
      • Mitochondrial lon peptidase 1 antibody
      • PIM1 antibody
      • Protease serine 15 antibody
      • PRSS15 antibody
      • Serine protease 15 antibody
      see all

    Images

    • Western blot - Anti-LONP1/Lon antibody (ab103809)
      Western blot - Anti-LONP1/Lon antibody (ab103809)
      Anti-LONP1/Lon antibody (ab103809) at 5 µg/ml + Human kidney tissue lysate at 50 µg

      Predicted band size: 106 kDa

    • Western blot - Anti-LONP1/Lon antibody (ab103809)
      Western blot - Anti-LONP1/Lon antibody (ab103809)
      Anti-LONP1/Lon antibody (ab103809) at 5 µg/ml + Mouse kidney tissue lysate at 50 µg

      Predicted band size: 106 kDa

    • Western blot - Anti-LONP1/Lon antibody (ab103809)
      Western blot - Anti-LONP1/Lon antibody (ab103809)
      Anti-LONP1/Lon antibody (ab103809) at 5 µg/ml + HepG2 cell lysate at 50 µg

      Predicted band size: 106 kDa

    • Western blot - Anti-LONP1/Lon antibody (ab103809)
      Western blot - Anti-LONP1/Lon antibody (ab103809)
      All lanes : Anti-LONP1/Lon antibody (ab103809) at 5 µg/ml

      Lane 1 : LONP1 transfected 293T cell lysate
      Lane 2 : Non-transfected 293T cell lysate

      Lysates/proteins at 25 µg per lane.

      Predicted band size: 106 kDa

    References

    This product has been referenced in:

    • Burstein SR  et al. In vitro and in vivo studies of the ALS-FTLD protein CHCHD10 reveal novel mitochondrial topology and protein interactions. Hum Mol Genet 27:160-177 (2018). Read more (PubMed: 29112723) »
    • Agnew T  et al. A Wars2 Mutant Mouse Model Displays OXPHOS Deficiencies and Activation of Tissue-Specific Stress Response Pathways. Cell Rep 25:3315-3328.e6 (2018). Read more (PubMed: 30566859) »
    See all 15 Publications for this product

    Publishing research using ab103809? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

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    1-6 of 6 Abreviews or Q&A

    Western blot abreview for Anti-LONP1/Lon antibody

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Tissue lysate - other (mitochondria isolated from placenta)
    Gel Running Conditions
    Reduced Denaturing (10%)
    Loading amount
    50 µg
    Specification
    mitochondria isolated from placenta
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
    Read More

    Dr. Polina Vishnyakova

    Verified customer

    Submitted Apr 01 2019

    Question

    mouse (untested)
    WB - sees multiple bands
    send discount code for testing ab76179 in mouse
    have peptide added to catalog

    Read More
    Answer

    Thank you for your phone call.

    I have requestedthat the immunogen peptide for ab76179 to be added to the catalog. I will let you know once I have more information.


    The information about the testing discount code for trying ab76179 in mouse can be found below as well as the instructions on how to redeem the code.

    Please let me know if you have further questions.

    DISCOUNT CODE: xxx
    Expiration date: xxx

    I am very pleased to hear you would like to accept our offer and test ab76179 in mouse. This code will give you: 1 free PRIMARY ANTIBODYbefore the expiration date. To redeem this offer, please submit an Abreview for mouseand 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.

    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: https://www.abcam.com/collaborationdiscount.

    Read More

    Question

    I am seeing no bands above 60kDa in my western blot; band is more like a smear; using same samples but different antibody to same target I am able to see expected band; samples are myosarcoma cell line extract

    Read More
    Answer

    Thank you for contacting us.


    I am sorry that this antibody did not perform as stated on the datasheet. If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact our accounting department so that we may gather the correct information needed for the refund. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used.

    Our accounting department can be contacted by email at us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website. Please refer to the credit note ID in any correspondence with our accounting department.

    The credit note ID is for your reference only and does not automatically guarantee the credit.

    I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expert advice.

    Read More

    Question

    Thank you for replying.
    The images I sent were including 4 pages, and I am sorry that it maygo withsomeproblem.
    Iseparate theoriginalfileinto 3images (including 2 independent experiments).Pleasehelpthis customercheck that. Regarding the tested sample,this customerworksinthehospital andclaims allthesamplescomes fromthepatients. All the fibroblast are from human newborns. Would you please help this customer check the data?
    Thank you very much. Best regards,

    Read More
    Answer

    Thank you very much for sending the images.

    We unfortunately have no other suggestions to make so we are happy to provide either free of charge replacement vial from different lot, refund or credit note. We have a new lot in stock so let me know if you are interested in getting a vial of this lot.

    Many thanks for your cooperation in this case. I will look forward to hearing from you soon.

    Read More

    Question

    Thank you for your reply.
    This customer has tried some modifications and sent back the results.
    Attached please find the data (4 pages).
    He has tried to:
    - Lyse sample with RIPA buffer.
    - Heat sample 95C for 10 mins. (previously he heated the samples in 70℃ for 10min is due to the protocol of preparing sample from the Invitrogen kit; http://tools.invitrogen.com/content/sfs/manuals/nupage_minigel_qrc.pdf)
    - Block with 5% BSA in TBST
    - 1Ab dilution(1:1000) with 5% BSA, 4C O/N
    - 2Ab dilution(1:5000) with 5% BSA in TBST at RT for 1Ab
    It is mentioned in a study (attached) that LonP1 are expressed in human blood, and he therefore would like see the clear signal like the reference showed. Unfortunately, multiple bands were still detected.
    Would you please help him check the data?
    Thank you very much.
    Best regards,

    Read More
    Answer

    Thank you for confirming the details.

    The image you have kindlyattached is the same images you have previously sent.Is there no improvements at all. Could you share any new image you have?

    Have you used lysates other than primaryfibroblasts? Are thesefibroblasts mouse or human?

    Looking forward to hearing from you soon.

    Read More

    Question

    LOT NUMBER GR43510-3 ORDER NUMBER 1007542 DESCRIPTION OF THE PROBLEM High background (LONP1 signal was so weakly.) SAMPLE - Species: human - What’s cell line or tissue: primary fibroblast cell (from the newborns) - Cell extract or Nuclear extract: cell extract - Purified protein or Recombinant protein: Whole cell extraction PRIMARY ANTIBODY - Species: rabbit polyclonal - Reacts against: human - At what dilution(s) have you tested this antibody: 1:1000 - What dilution buffer was used: 5% non-fat-milk in TBST - Incubation time: 1hr - Incubation temperature: room temperature - What washing steps were done: 15min x 1 + 5min x 3 DETECTION METHOD ECl+ ANTIBODY STORAGE CONDITIONS -20 SAMPLE PREPARATION - What lysis buffer was used: 20mM potassium phosphate buffer + sonication - What protease inhibitors were used: cocktail protease inhibitor - What loading buffer was used: NuPAGE LDS sample buffer (invitrogen:Cat no.NP0007) - Phosphatase inhibitors - Did you heat the samples: temperature and time: 70℃ for 10min AMOUNT OF PROTEIN LOADED 30ug ELECTROPHORESIS/GEL CONDITIONS - Reducing or non reducing gel: Reducing - Reducing agent: 2-ME - Gel percentage : 4-12% TRANSFER AND BLOCKING CONDITIONS - Transfer conditions: (Type of membrane, Protein transfer verified): PVDF, Coomassie blue (Invitrogen iBlot dry blotting system Cat no. IB10001) Blocking conditions - Buffer: 5% non-fat-milk in TBST - Blocking agent: milk, BSA, serum, what percentage: 5% - Incubation time: 1hr - Incubation temperature: room temperature SECONDARY ANTIBODY - Species: goat - Reacts against: rabbit - At what dilution(s) have you tested this antibody: 1:5000 - Incubation time: 1hr - Wash steps: 15min x 1 + 5min x 3 - Fluorochrome or enzyme conjugate: enzyme conjugate - Do you know whether the problems you are experiencing come from the secondary? HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 1 HAVE YOU RUN A "NO PRIMARY" CONTROL? No

    Read More
    Answer

    Thank you for your enquiry regarding ab103809 and for taking the time to provide some useful details of the experiments. I am very sorry to hear that you are having problems with this antibody. Additionally, thank you for supplying an image, as this has been very beneficial in understanding your concerns.
    The protocol looks fine to me however I would like to make few comments/suggestions that might help to improve your results and that you could consider trying:
    The 100 kDa band presented on the online datasheet is with tissue lysates of human kidney, mouse and cell lysates of HepG2 cells. After reviewing the image you have kindly sent it looks like the ab is showing multiple bands. I would recommend troubleshooting the protocol further. My suggestions would be
    - Not sure if phosphate buffer was used a lysis buffer? We recommend using NP40 buffer.
    - When doing wb we heated the sample for 10 minutes at 95-100 C. Could you tty the same?
    - We normally use 5% BSA in TBST (2 hrs 4C) for antibody dilution and blocking? I would recommend trying this.
    - Try the above mentioned tissue lysates as positive control.
    I am sure changing these steps will help to improve results. Should the results do not improve please do not hesitate to contact me.

    Read More

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