Overview

  • Product name

    Anti-Ferritin Heavy Chain antibody
    See all Ferritin Heavy Chain primary antibodies
  • Description

    Rabbit polyclonal to Ferritin Heavy Chain
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Dog, Human
    Predicted to work with: Sheep, Goat, Cow, Cat, Orangutan
  • Immunogen

    Synthetic peptide corresponding to Human Ferritin Heavy Chain aa 1-100 (N terminal) conjugated to keyhole limpet haemocyanin.
    Database link: P02794
    (Peptide available as ab90631)

  • Positive control

    • Recombinant Human Ferritin Heavy Chain protein (ab78877) can be used as a positive control in WB. This antibody gave a positive signal in Human Liver Tissue Lysate. IF/ICC: MCF7 cell line. IHC-P: FFPE human normal liver tissue sections.

Properties

Applications

Our Abpromise guarantee covers the use of ab65080 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 µg/ml. Detects a band of approximately 21 kDa (predicted molecular weight: 21 kDa).
IHC-P Use a concentration of 5 µg/ml.
ICC/IF Use a concentration of 5 µg/ml.

Target

Images

  • Anti-Ferritin Heavy Chain antibody (ab65080) at 1 µg/ml + Human liver tissue lysate - total protein (ab29889) at 10 µg

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 21 kDa
    Observed band size: 21 kDa
    Additional bands at: 24 kDa (possible post-translational modification)


    Exposure time: 3 minutes


    Ferritin Heavy Chain contains a number of potential phosphorylation sites (SwissProt data) which may explain it running at a slightly higher than predicted molecular weight (24 kDa).
  • ICC/IF image of ab65080 stained MCF-7 cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab65080, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 Goat anti-Rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
  • IHC image of ab65080 staining in Human Normal Liver formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab65080, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
  • All lanes : Anti-Ferritin Heavy Chain antibody (ab65080) at 1 µg/ml

    Lane 1 : Recombinant Human Ferritin Heavy Chain protein (ab78877) at 0.1 µg
    Lane 2 : Recombinant Human Ferritin Heavy Chain protein (ab78877) at 0.01 µg

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 21 kDa


    Exposure time: 30 seconds

References

This product has been referenced in:

  • Kong Z  et al. Artesunate alleviates liver fibrosis by regulating ferroptosis signaling pathway. Biomed Pharmacother 109:2043-2053 (2019). Read more (PubMed: 30551460) »
  • Cui R  et al. Iron overload by transferrin receptor protein 1 regulation plays an important role in palmitate-induced insulin resistance in human skeletal muscle cells. FASEB J 33:1771-1786 (2019). Read more (PubMed: 30207798) »
See all 29 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (human breast cancer cells)
Total protein in input
1e+007 cells
Immuno-precipitation step
Protein A/G
Specification
human breast cancer cells

Abcam user community

Verified customer

Submitted Aug 22 2018

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Mouse Retina)
Permeabilization
No
Specification
Mouse Retina
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Aug 03 2016

Application
Flow Cytometry
Sample
Human Cell (Differentiated hNSCs)
Permeabilization
Yes - 0.25% Triton X-100 in DPBS
Gating Strategy
Undifferentiated Stem Cells (white)
Specification
Differentiated hNSCs
Preparation
Cell harvesting/tissue preparation method: Accutase
Sample buffer: PBS
Fixation
Paraformaldehyde

Abcam user community

Verified customer

Submitted Sep 09 2015

Answer

Thank you for contacting Abcam.

The antibody is covered under our Abpromise for six months and is guaranteed to work in western blot on mouse and human samples . If we cannot resolve the issue you are having with the antibody then I would be happy to either send a replacement antibody or to process a refund.

I know that you said the other 4 antibodies you ordered worked well for you, but would you be able to provide a little more information on the samples and protocols you were using with ab84036.

1 - What sample type are you using?

2 - Are you seeing any bands, no bands etc?

3 - How much sample are you loading?

4 - What type of blocking agent are you using?

5 - What primary antibody concentrations have you tried and how long do you incubate for?

As I mentioned before, if you are using it on mouse or human samples and we cannot resolve the issue you are having, I will be happy to either replace or refund the cost of the antibody.

I look forward to your reply.

Read More

Answer

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement of ab65080.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

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