Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Ferritin Light Chain antibody [EPR5260] (ab109373)

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR5260] to Ferritin Light Chain
  • Suitable for: WB, Flow Cyt
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-Ferritin Light Chain antibody [EPR5260]
    See all Ferritin Light Chain primary antibodies

  • Description

    Rabbit monoclonal [EPR5260] to Ferritin Light Chain

  • Host species

    Rabbit

  • Specificity

    ab109373 recognizes the light chain of Ferritin only.

  • Tested applications

    Suitable for: WB, Flow Cytmore details
    Unsuitable for: ICC or IHC-P

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide within Human Ferritin Light Chain aa 150-250 (C terminal). The exact sequence is proprietary.

  • Positive control

    • Human fetal liver, Hap1, HepG2 and HeLa cell lysates. Flow Cyt: HeLa cells. IHC-P: Mouse brain tissue.

  • General notes

     

     

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Applications

Our Abpromise guarantee covers the use of ab109373 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 1/10000 - 1/50000. Detects a band of approximately 19 kDa (predicted molecular weight: 20 kDa).
Flow Cyt 1/100 - 1/500.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 
  • Application notes
    Is unsuitable for ICC or IHC-P.

    • Target

    • Function

      Stores iron in a soluble, non-toxic, readily available form. Important for iron homeostasis. Iron is taken up in the ferrous form and deposited as ferric hydroxides after oxidation. Also plays a role in delivery of iron to cells. Mediates iron uptake in capsule cells of the developing kidney.

    • Involvement in disease

      Defects in FTL are the cause of hereditary hyperferritinemia-cataract syndrome (HHCS) [MIM:600886]. It is an autosomal dominant disease characterized by early-onset bilateral cataract. Affected patients have elevated level of circulating ferritin. HHCS is caused by mutations in the iron responsive element (IRE) of the FTL gene.
      Defects in FTL are the cause of neurodegeneration with brain iron accumulation type 3 (NBIA3) [MIM:606159]; also known as adult-onset basal ganglia disease. It is a movement disorder with heterogeneous presentations starting in the fourth to sixth decade. It is characterized by a variety of neurological signs including parkinsonism, ataxia, corticospinal signs, mild nonprogressive cognitive deficit and episodic psychosis. It is linked with decreased serum ferritin levels.

    • Sequence similarities

      Belongs to the ferritin family.
      Contains 1 ferritin-like diiron domain.
    • Information by UniProt

    • Database links

      see all

    • Alternative names

      • Ferritin L chain antibody
      • Ferritin L subunit antibody
      • Ferritin light chain antibody
      • Ferritin light polypeptide antibody
      • ferritin light polypeptide like 3 antibody
      • FRIL_HUMAN antibody
      • FTL antibody
      • LFTD antibody
      • NBIA 3 antibody
      • NBIA3 antibody
      see all

    Images

    • Western blot - Anti-Ferritin Light Chain antibody [EPR5260] (ab109373)
      Western blot - Anti-Ferritin Light Chain antibody [EPR5260] (ab109373)
      All lanes : Anti-Ferritin Light Chain antibody [EPR5260] (ab109373) at 1/2000 dilution

      Lane 1 : Wild-type HeLa cell lysate
      Lane 2 : Ferritin knockout HeLa cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 20 kDa



      Lanes 1-2: Merged signal (red and green). Green - ab109373 observed at 20 kDa. Red - loading control ab8245 observed at 37 kDa.

       ab109373 Anti-FTL was shown to specifically react with Ferritin in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265533 (knockout cell lysate ab256926) was used. Wild-type and FTL knockout samples were subjected to SDS-PAGE. ab109373 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 2000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

       

    • Western blot - Anti-Ferritin Light Chain antibody [EPR5260] (ab109373)
      Western blot - Anti-Ferritin Light Chain antibody [EPR5260] (ab109373)
      All lanes : Anti-Ferritin Light Chain antibody [EPR5260] (ab109373) at 1/10000 dilution

      Lane 1 : Wild-type HAP1 whole cell lysate
      Lane 2 : FTL (Ferritin Light Chain) knockout HAP1 whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 20 kDa



      Lanes 1 - 2: Merged signal (red and green). Green - ab109373 observed at 20 kDa. Red - loading control, ab9484, observed at 37 kDa.

      ab109373 was shown to recognize Ferritin Light Chain in wild-type HAP1 cells as signal was lost at the expected MW in FTL (Ferritin Light Chain) knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and FTL (Ferritin Light Chain) knockout samples were subjected to SDS-PAGE. Ab109373 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/10000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ferritin Light Chain antibody [EPR5260] (ab109373)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ferritin Light Chain antibody [EPR5260] (ab109373)Li W et al., 10(1):e0117435. Fig 5 (A-B). doi:10.1371/journal.pone.0117435. Reproduced under the Creative Commons license https://creativecommons.org/licenses/by/4.0/

      Immunohistochemical analysis of wild type and ftl knock out mouse brain tissue labeling Ferritin Light Chain. Positive staining of Ferratin Light Chain is seen in wild type mouse brain (A) and no staining is seen in the ftl knock out brain tissue (B).

    • Western blot - Anti-Ferritin Light Chain antibody [EPR5260] (ab109373)
      Western blot - Anti-Ferritin Light Chain antibody [EPR5260] (ab109373)
      All lanes : Anti-Ferritin Light Chain antibody [EPR5260] (ab109373) at 1/10000 dilution

      Lane 1 : Fetal liver lysate
      Lane 2 : HepG2 cell lysate
      Lane 3 : HeLa cell lysate

      Lysates/proteins at 10 µg per lane.

      Predicted band size: 20 kDa
      Observed band size: 19 kDa
      why is the actual band size different from the predicted?

    • Flow Cytometry - Anti-Ferritin Light Chain antibody [EPR5260] (ab109373)
      Flow Cytometry - Anti-Ferritin Light Chain antibody [EPR5260] (ab109373)
      Flow cytometric analysis of permeabilized HeLa cells using ab109373 at a dilution of 1/100 (red) or a rabbit IgG (negative) (green).
    • Anti-Ferritin Light Chain antibody [EPR5260] (ab109373)
      Anti-Ferritin Light Chain antibody [EPR5260] (ab109373)

    References (4)

    Publishing research using ab109373? Please let us know so that we can cite the reference in this datasheet.

    ab109373 has been referenced in 4 publications.

    • Wang K  et al. Correction of Anemia in Chronic Kidney Disease With Angelica sinensis Polysaccharide via Restoring EPO Production and Improving Iron Availability. Front Pharmacol 9:803 (2018). PubMed: 30108502
    • Khiroya H  et al. IRP2 as a potential modulator of cell proliferation, apoptosis and prognosis in nonsmall cell lung cancer. Eur Respir J 49:N/A (2017). PubMed: 28404645
    • Mertens C  et al. Intracellular Iron Chelation Modulates the Macrophage Iron Phenotype with Consequences on Tumor Progression. PLoS One 11:e0166164 (2016). Flow Cyt ; Human . PubMed: 27806101
    • Li W  et al. Systemic and cerebral iron homeostasis in ferritin knock-out mice. PLoS One 10:e0117435 (2015). WB, IHC-P ; Mouse . PubMed: 25629408

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