Anti-Frataxin antibody [EPR21840] - BSA and Azide free (ab236463)

Rabbit Recombinant Monoclonal Frataxin antibody. Carrier free. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Mouse, Human, Recombinant fragment - Human, Rat samples.

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Images

Immunoprecipitation - Anti-Frataxin antibody [EPR21840] - BSA and Azide free (AB236463), expandable thumbnail

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: PBS
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IP	WB	ICC/IF	Flow Cyt (Intra)	IHC-P
Human	Expected	Expected	Tested	Tested	Tested
Mouse	Tested	Expected	Expected	Expected	Expected
Rat	Predicted	Expected	Predicted	Predicted	Predicted
Recombinant fragment - Human	Not recommended	Tested	Not recommended	Not recommended	Not recommended

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Human	Dilution info Use at an assay dependent concentration.	Notes -

Predicted
Predicted

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Recombinant fragment - Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Recombinant fragment - Human	Dilution info -	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat, Human	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse	Dilution info Use at an assay dependent concentration.	Notes -

Predicted
Predicted

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Recombinant fragment - Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse	Dilution info Use at an assay dependent concentration.	Notes -

Predicted
Predicted

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Recombinant fragment - Human	Dilution info -	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info -	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Mouse	Dilution info Use at an assay dependent concentration.	Notes -

Predicted
Predicted

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Recombinant fragment - Human	Dilution info -	Notes -

Associated Products

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Target data

See full target information FXN

Function

Frataxin mature form. Functions as an activator of persulfide transfer to the scaffoding protein ISCU as component of the core iron-sulfur cluster (ISC) assembly complex and participates to the [2Fe-2S] cluster assembly (PubMed:12785837, PubMed:24971490). Accelerates sulfur transfer from NFS1 persulfide intermediate to ISCU and to small thiols such as L-cysteine and glutathione leading to persulfuration of these thiols and ultimately sulfide release (PubMed:24971490). Binds ferrous ion and is released from FXN upon the addition of both L-cysteine and reduced FDX2 during [2Fe-2S] cluster assembly (PubMed:29576242). The core iron-sulfur cluster (ISC) assembly complex is involved in the de novo synthesis of a [2Fe-2S] cluster, the first step of the mitochondrial iron-sulfur protein biogenesis. This process is initiated by the cysteine desulfurase complex (NFS1:LYRM4:NDUFAB1) that produces persulfide which is delivered on the scaffold protein ISCU in a FXN-dependent manner. Then this complex is stabilized by FDX2 which provides reducing equivalents to accomplish the [2Fe-2S] cluster assembly. Finally, the [2Fe-2S] cluster is transferred from ISCU to chaperone proteins, including HSCB, HSPA9 and GLRX5 (By similarity). 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 (PubMed:15641778). 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 (PubMed:11823441, PubMed:12755598). May function as an iron chaperone protein that protects the aconitase [4Fe-4S]2+ cluster from disassembly and promotes enzyme reactivation (PubMed:15247478). May play a role as a high affinity iron binding partner for FECH that is capable of both delivering iron to ferrochelatase and mediating the terminal step in mitochondrial heme biosynthesis (PubMed:15123683, PubMed:16239244). Extramitochondrial frataxin. Modulates the RNA-binding activity of ACO1 (PubMed:20053667). May be involved in the cytoplasmic iron-sulfur protein biogenesis (PubMed:16091420). May contribute to oxidative stress resistance and overall cell survival (PubMed:16608849).

Alternative names

FRDA, X25, FXN, Friedreich ataxia protein, Fxn

Recommended products

Rabbit Recombinant Monoclonal Frataxin antibody. Carrier free. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra), IHC-P and reacts with Mouse, Human, Recombinant fragment - Human, Rat samples.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: PBS
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Carrier free: Yes
Clone number: EPR21840
Purification technique: Affinity purification Protein A
Specificity: We suggest optimizing experimental protocols (increasing lysate amount, using lower dilution or higher sensitivity ECL substrate) to improve results.
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: +4°C
Storage information: Do Not Freeze

Notes

ab236463 is the carrier-free version of Anti-Frataxin antibody [EPR21840] ab219414.

Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

This product is compatible with the Maxpar^® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar^® is a trademark of Fluidigm Canada Inc.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

Frataxin often known by the alternate name FXN is a mitochondrial protein with a mass of approximately 21000 Dalton. It is expressed mainly in tissues with high energy demands like the heart liver and pancreas. Frataxin plays an important role in iron-sulfur cluster assembly which is essential for various cellular processes. The protein is a part of mitochondria where it regulates iron homeostasis and prevents oxidative damage by minimizing iron-induced free radical generation.

Biological function summary

Several cellular processes depend on the correct function of this protein. Frataxin assists in forming iron-sulfur clusters acting within a multiprotein complex in the mitochondria. The complex includes proteins such as ISCU which are involved in the assembly and repair of iron-sulfur clusters. These clusters are necessary for supporting mitochondrial electron transport and other fundamental metabolic pathways that require iron-sulfur dependencies.

Pathways

Frataxin's involvement extensively affects the mitochondrial respiratory chain and the mitochondrial biogenesis process. It plays a role in the electron transport chain by stabilizing iron-sulfur-containing complexes. NAB is one associated protein that interacts closely within these pathways sharing a connection through iron-sulfur cluster transportation and assembly systems. Efficient function of these pathways ensures a proper energetic output of cells.

Associated diseases and disorders

Frataxin mutations are directly linked to Friedreich's ataxia a neurodegenerative disease causing progressive damage to the nervous system. The deficiency or dysfunction in frataxin causes accumulation of iron in mitochondria leading to increased oxidative stress. Another related disorder includes heart disease which emerges due to the same oxidative stress pathway. Proteins such as Nfs1 are also involved sharing the responsibility with frataxin in scavenging excess iron protecting against related tissue damage.

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11 product images

Immunoprecipitation - Anti-Frataxin antibody [EPR21840] - BSA and Azide free (ab236463)
Frataxin was immunoprecipitated from 0.35 mg Neuro-2a (mouse neuroblastoma cell line) whole cell lysate with Anti-Frataxin antibody [EPR21840] ab219414 at 1/30 dilution. Western blot was performed from the immunoprecipitate using Anti-Frataxin antibody [EPR21840] ab219414 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (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: Anti-Frataxin antibody [EPR21840] ab219414 IP in Neuro-2a whole cell lysate (+).
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-Frataxin antibody [EPR21840] ab219414 in Neuro-2a whole cell lysate (-).

Blocking/Dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 30 seconds.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Frataxin antibody [EPR21840] ab219414).
All lanes: Immunoprecipitation - Anti-Frataxin antibody [EPR21840] (Anti-Frataxin antibody [EPR21840] ab219414)
Predicted band size: 23 kDa
Observed band size: 14 kDa
Immunoprecipitation - Anti-Frataxin antibody [EPR21840] - BSA and Azide free (ab236463)
Frataxin was immunoprecipitated from 0.35 mg HCT 116 (human colorectal carcinoma epithelial cell) whole cell lysate with Anti-Frataxin antibody [EPR21840] ab219414 at 1/30 dilution. Western blot was performed from the immunoprecipitate using Anti-Frataxin antibody [EPR21840] ab219414 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (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: Anti-Frataxin antibody [EPR21840] ab219414 IP in HCT 116 whole cell lysate (+).
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-Frataxin antibody [EPR21840] ab219414 in HCT 116 whole cell lysate (-).

Blocking/Dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 5 seconds.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Frataxin antibody [EPR21840] ab219414).
All lanes: Immunoprecipitation - Anti-Frataxin antibody [EPR21840] (Anti-Frataxin antibody [EPR21840] ab219414)
Predicted band size: 13 kDa, 23 kDa
Observed band size: 17 kDa
Immunocytochemistry/ Immunofluorescence - Anti-Frataxin antibody [EPR21840] - BSA and Azide free (ab236463)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HCT 116 (human colorectal carcinoma epithelial cell) cells labeling Frataxin with Anti-Frataxin antibody [EPR21840] ab219414 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (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 Anti-COX IV antibody [mAbcam33985] - Mitochondrial Marker ab33985 Anti-COX IV antibody - Mitochondrial Marker at a 1/1000 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) (orange). The nuclear counter stain is DAPI (blue).
The negative control is the secondary antibody only.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Frataxin antibody [EPR21840] ab219414).
Flow Cytometry (Intracellular) - Anti-Frataxin antibody [EPR21840] - BSA and Azide free (ab236463)
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeabilized HCT 116 (human colorectal carcinoma epithelial cell) cell line labeling Frataxin with Anti-Frataxin antibody [EPR21840] ab219414 at 1/60 (red) compared with a rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077), at 1/2000 dilution was used as the secondary antibody.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Frataxin antibody [EPR21840] ab219414).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Frataxin antibody [EPR21840] - BSA and Azide free (ab236463)
Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling Frataxin with Anti-Frataxin antibody [EPR21840] 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 Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer pH 9.0).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Frataxin antibody [EPR21840] ab219414).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Frataxin antibody [EPR21840] - BSA and Azide free (ab236463)
Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling Frataxin with Anti-Frataxin antibody [EPR21840] 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 Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer pH 9.0).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Frataxin antibody [EPR21840] ab219414).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Frataxin antibody [EPR21840] - BSA and Azide free (ab236463)
Immunohistochemical analysis of paraffin-embedded human liver tissue labeling Frataxin with Anti-Frataxin antibody [EPR21840] 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 Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer pH 9.0).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Frataxin antibody [EPR21840] ab219414).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Frataxin antibody [EPR21840] - BSA and Azide free (ab236463)
Immunohistochemical analysis of paraffin-embedded human testis tissue labeling Frataxin with Anti-Frataxin antibody [EPR21840] 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 Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer pH 9.0).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Frataxin antibody [EPR21840] ab219414).
Immunocytochemistry/ Immunofluorescence - Anti-Frataxin antibody [EPR21840] - BSA and Azide free (ab236463)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Neuro-2a (mouse neuroblastoma cell line) cells labeling Frataxin with Anti-Frataxin antibody [EPR21840] ab219414 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (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 Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Frataxin antibody [EPR21840] ab219414).
Western blot - Anti-Frataxin antibody [EPR21840] - BSA and Azide free (ab236463)
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Frataxin antibody [EPR21840] ab219414).

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

Anti-6X His tag® antibody [EPR20547] - ChIP Grade ab213204 was used as laoding control.
All lanes: Western blot - Anti-Frataxin antibody [EPR21840] (Anti-Frataxin antibody [EPR21840] ab219414) at 1/1000 dilution
All lanes: His tagged Human FXN recombinant protein (aa 82-210) at 10 ng
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 23 kDa
Observed band size: 16 kDa
Exposure time: 3s
Western blot - Anti-Frataxin antibody [EPR21840] - BSA and Azide free (ab236463)
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (Anti-Frataxin antibody [EPR21840] ab219414).
Blocking and diluting buffer and concentration: 5% NFDM/TBST.
This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.
Anti-Frataxin antibody [EPR21840] ab219414 is not suitable for testing precursor form, we recommend Anti-Frataxin antibody [EPR6107] ab124680 as an alternative for precursor form testing.
For different forms of frataxin, you can refer to PMID: 17468497, PMID: 31279523, PMID: 17468497 etc.
Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 was used as a loading control.
All lanes: Western blot - Anti-Frataxin antibody [EPR21840] (Anti-Frataxin antibody [EPR21840] ab219414) at 1/1000 dilution
Lane 1: Human liver tissue lysate at 20 µg
Lane 2: Human hippocampus tissue at 20 µg
Lane 3: HCT116 (human colorectal carcinoma epithelial cell) whole cell lysate at 20 µg
Lane 4: Huh7 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg
Lane 5: SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Developed using the ECL technique.
Predicted band size: 23 kDa
Observed band size: 14,17 kDa
Exposure time: 60s