Recombinant Anti-Frataxin antibody [EPR21840] (ab219414)

Product name

Anti-Frataxin antibody [EPR21840]
See all Frataxin primary antibodies
Description

Rabbit monoclonal [EPR21840] to Frataxin
Host species

Rabbit
Tested applications

Suitable for: WB, IHC-P, IP, Flow Cyt, ICC/IFmore details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Recombinant fragment within Human Frataxin aa 50 to the C-terminus. The exact sequence is proprietary.
Database link: Q16595
Positive control
- WB: HCT 116, Neuro-2a, C6 and PC-12 whole cell lysates. IHC-P: Human testis and liver tissue; Mouse and rat kidney tissue. ICC/IF: HCT 116 and Neuro-2a cells. Flow Cyt: HCT 116 cells. IP: HCT 116 and Neuro-2a cells.
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.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 0.05% BSA, 40% Glycerol (glycerin, glycerine)
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR21840
Isotype

IgG
Research areas

Alternative Versions
- Anti-Frataxin antibody [EPR21840] - BSA and Azide free (ab236463)
Compatible Secondaries
Conjugation kits
- Biotinylation Kit / Biotin Conjugation Kit (Fast, Type A) - Lightning-Link® (ab201795)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant Human Frataxin protein (ab110353)
Related Products

Our Abpromise guarantee covers the use of ab219414 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		1/1000. Detects a band of approximately 14 kDa (predicted molecular weight: 23 kDa).
IHC-P		1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP		1/30.
Flow Cyt		1/60.
ICC/IF		1/50.

Function

Promotes the biosynthesis of heme and assembly and repair of iron-sulfur clusters by delivering Fe(2+) to proteins involved in these pathways. May play a role in the protection against iron-catalyzed oxidative stress through its ability to catalyze the oxidation of Fe(2+) to Fe(3+); the oligomeric form but not the monomeric form has in vitro ferroxidase activity. May be able to store large amounts of iron in the form of a ferrihydrite mineral by oligomerization; however, the physiological relevance is unsure as reports are conflicting and the function has only been shown using heterologous overexpression systems. Modulates the RNA-binding activity of ACO1.
Tissue specificity

Expressed in the heart, peripheral blood lymphocytes and dermal fibroblasts.
Involvement in disease

Defects in FXN are the cause of Friedreich ataxia (FRDA) [MIM:229300]. FRDA is an autosomal recessive, progressive degenerative disease characterized by neurodegeneration and cardiomyopathy it is the most common inherited ataxia. The disorder is usually manifest before adolescence and is generally characterized by incoordination of limb movements, dysarthria, nystagmus, diminished or absent tendon reflexes, Babinski sign, impairment of position and vibratory senses, scoliosis, pes cavus, and hammer toe. In most patients, FRDA is due to GAA triplet repeat expansions in the first intron of the frataxin gene. But in some cases the disease is due to mutations in the coding region.
Sequence similarities

Belongs to the frataxin family.
Post-translational
modifications

Processed in two steps by mitochondrial processing peptidase (MPP). MPP first cleaves the precursor to intermediate form and subsequently converts the intermediate to yield frataxin mature form (frataxin(81-210)) which is the predominant form. The additional forms, frataxin(56-210) and frataxin(78-210), seem to be produced when the normal maturation process is impaired; their physiological relevance is unsure.
Cellular localization

Cytoplasm. Mitochondrion. PubMed:18725397 reports localization exclusively in mitochondria.
Target information above from: UniProt accession Q16595 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 2395 Human
- Entrez Gene: 14297 Mouse
- Entrez Gene: 499335 Rat
- Omim: 606829 Human
- SwissProt: Q16595 Human
- SwissProt: O35943 Mouse
- SwissProt: D3ZYW7 Rat
- Unigene: 20685 Human
- Unigene: 7319 Mouse
- Unigene: 13133 Rat
see all
Alternative names
- CyaY antibody
- d-FXN antibody
- FA antibody
- FARR antibody
- Frataxin mature form antibody
- Frataxin(81-210) antibody
- FRDA antibody
- FRDA_HUMAN antibody
- Friedreich ataxia protein antibody
- Fxn antibody
- i-FXN antibody
- m56-FXN antibody
- m78-FXN antibody
- m81-FXN antibody
- MGC57199 antibody
- X25 antibody
see all

Western blot - Anti-Frataxin antibody [EPR21840] (ab219414)

All lanes : Anti-Frataxin antibody [EPR21840] (ab219414) at 1/1000 dilution

Lane 1 : HCT 116 (human colorectal carcinoma cell line) whole cell lysate
Lane 2 : Neuro-2a (mouse neuroblastoma cell line) whole cell lysate
Lane 3 : C6 (rat glial tumor glial cell) whole cell lysate
Lane 4 : PC-12 (rat adrenal gland pheochromocytoma cell line) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
Lanes 1-2 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
Lanes 3-4 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 23 kDa
Observed band size: 14 kDa
why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blockingand dilution buffer: 5% NFDM/TBST .
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Frataxin antibody [EPR21840] (ab219414)

Immunohistochemical analysis of paraffin-embedded human testis tissue labeling Frataxin with ab219414 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Granular cytoplasmic staining in human testis (PMID: 18725397; PMID: 26035392) is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Perform heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer pH 9.0).
Immunocytochemistry/ Immunofluorescence - Anti-Frataxin antibody [EPR21840] (ab219414)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HCT 116 (human colorectal carcinoma epithelial cell) cells labeling Frataxin with ab219414 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing mitochondrial staining in HCT 116 cell line.

Counterstained with ab33985 Anti-COX IV antibody - Mitochondrial Marker at a 1/1000 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) (orange). The nuclear counter stain is DAPI (blue).

The negative control is the secondary antibody only.
Immunoprecipitation - Anti-Frataxin antibody [EPR21840] (ab219414)

Frataxin was immunoprecipitated from 0.35 mg Neuro-2a (mouse neuroblastoma cell line) whole cell lysate with ab219414 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab219414 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

Lane 1: Neuro-2a whole cell lysate 10 µg (Input).
Lane 2: ab219414 IP in Neuro-2a whole cell lysate (+).
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab219414 in Neuro-2a whole cell lysate (-).

Blocking/Dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 30 seconds.
Immunoprecipitation - Anti-Frataxin antibody [EPR21840] (ab219414)

Frataxin was immunoprecipitated from 0.35 mg HCT 116 (human colorectal carcinoma epithelial cell) whole cell lysate with ab219414 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab219414 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

Lane 1: HCT 116 whole cell lysate 10 µg (Input).
Lane 2: ab219414 IP in HCT 116 whole cell lysate (+).
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab219414 in HCT 116 whole cell lysate (-).

Blocking/Dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 5 seconds.
Flow Cytometry - Anti-Frataxin antibody [EPR21840] (ab219414)

Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HCT 116 (human colorectal carcinoma epithelial cell) cell line labeling Frataxin with ab219414 at 1/60 (red) compared with a rabbit monoclonal IgG (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Frataxin antibody [EPR21840] (ab219414)

Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling Frataxin with ab219414 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Granular cytoplasmic staining in rat kidney (PMID: 18725397; PMID: 26035392) is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Perform heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer pH 9.0).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Frataxin antibody [EPR21840] (ab219414)

Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling Frataxin with ab219414 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Granular cytoplasmic staining in mouse kidney (PMID: 18725397; PMID: 26035392) is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Perform heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer pH 9.0).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Frataxin antibody [EPR21840] (ab219414)

Immunohistochemical analysis of paraffin-embedded human liver tissue labeling Frataxin with ab219414 at 1/500 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Granular cytoplasmic staining in human liver (PMID: 18725397; PMID: 26035392) is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

Perform heat-mediated antigen retrieval using ab93684 (Tris/EDTA buffer pH 9.0).
Immunocytochemistry/ Immunofluorescence - Anti-Frataxin antibody [EPR21840] (ab219414)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Neuro-2a (mouse neuroblastoma cell line) cells labeling Frataxin with ab219414 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing cytoplasmic staining in the Neuro-2a cell line.

Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) at a 1/200 dilution (red). The nuclear counter stain is DAPI (blue).

The negative control is the secondary antibody only.
Anti-Frataxin antibody [EPR21840] (ab219414)

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

Datasheet

Publishing research using ab219414? Please let us know so that we can cite the reference in this datasheet.

ab219414 has been referenced in 1 publication.

Du J et al. Identification of Frataxin as a regulator of ferroptosis. Redox Biol 32:101483 (2020). PubMed: 32169822

Key features and details

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Conjugation kits

Isotype control

Recombinant Protein

Related Products

Applications

Target

Function

Tissue specificity

Involvement in disease

Sequence similarities

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

Certificate of Compliance

References (1)

Customer reviews and Q&As

Post-translational
modifications