Recombinant Anti-Frataxin antibody [EPR21840] (ab219414)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR21840] to Frataxin
- Suitable for: WB, IHC-P, IP, ICC/IF, Flow Cyt (Intra)
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
-
Product name
Anti-Frataxin antibody [EPR21840]
See all Frataxin primary antibodies -
Description
Rabbit monoclonal [EPR21840] to Frataxin -
Host species
Rabbit -
Specificity
We suggest optimizing experimental protocols (increasing lysate amount, using lower dilution or higher sensitivity ECL substrate) to improve results.
-
Tested applications
Suitable for: WB, IHC-P, IP, ICC/IF, Flow Cyt (Intra)more details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
-
Positive control
- WB: HCT 116, Neuro-2a, C6, PC-12, human liver and human hippocampus tissue lysates, HCT116, Huh7 and SH-SY5Y 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 (intra): 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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
-
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
Associated products
-
Alternative Versions
-
Compatible Secondaries
-
Conjugation kits
-
Isotype control
-
Related Products
Applications
The Abpromise guarantee
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.
|
|
ICC/IF |
1/50.
|
|
Flow Cyt (Intra) |
1/60.
|
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. |
ICC/IF
1/50. |
Flow Cyt (Intra)
1/60. |
Target
-
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
modificationsProcessed 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. - 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
see all -
Alternative names
- CyaY antibody
- d-FXN antibody
- FA antibody
see all
Images
-
All lanes : ab219414 and ab124680 at 1/1000 dilution
Lane 1 : Human liver tissue lysate
Lane 2 : Human hippocampus tissue lysate
Lane 3 : HCT116 (human colorectal carcinoma epithelial cell) whole cell lysate
Lane 4 : Huh7 (human hepatocellular carcinoma epithelial cell) whole cell lysate
Lane 5 : SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 23 kDa
Observed band size: 14,17 kDa why is the actual band size different from the predicted?
Exposure time: 60 secondsBlocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
This blot was developed using a high sensitivity ECL substrate.
ab219414 is not suitable for testing precursor form, we recommend ab124680 as an alternative for precursor form testing.
For different forms of frataxin, you can refer to PMID: 17468497, PMID: 31279523, PMID: 17468497 etc.
ab181602 was used as a loading control.
-
Anti-Frataxin antibody [EPR21840] (ab219414) at 1/1000 dilution + His tagged Human FXN recombinant protein (aa 82-210), 10ng
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 23 kDa
Observed band size: 16 kDa why is the actual band size different from the predicted?
Exposure time: 3 secondsBlocking and diluting buffer and concentration: 5% NFDM/TBST.
ab213204 was used as laoding control.
-
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 minutesBlockingand dilution buffer: 5% NFDM/TBST .
-
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).
-
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.
-
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. -
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. -
Intracellular 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.
-
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).
-
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).
-
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).
-
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.
Protocols
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.
Datasheets and documents
-
SDS download
-
Datasheet download
Certificate of Compliance
References (2)
ab219414 has been referenced in 2 publications.
- Medina-Carbonero M et al. Mice harboring the FXN I151F pathological point mutation present decreased frataxin levels, a Friedreich ataxia-like phenotype, and mitochondrial alterations. Cell Mol Life Sci 79:74 (2022). PubMed: 35038030
- Du J et al. Identification of Frataxin as a regulator of ferroptosis. Redox Biol 32:101483 (2020). PubMed: 32169822