Anti-Frataxin antibody [EPR21840]

• ICC/IF

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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.

• IHC-P

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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).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Frataxin antibody [EPR21840] (AB219414)

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.

• IHC-P

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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).

• IP

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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.

All lanes:

Immunoprecipitation - Anti-Frataxin antibody [EPR21840] (ab219414)

Predicted band size: 13 kDa,23 kDa

Observed band size: 17 kDa

false

• IHC-P

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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).

• ICC/IF

Unknown

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.

• IHC-P

Unknown

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).

• IP

Unknown

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.

All lanes:

Immunoprecipitation - Anti-Frataxin antibody [EPR21840] (ab219414)

Predicted band size: 23 kDa

Observed band size: 14 kDa

false

• WB

Supplier Data

Western blot - 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.

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.

All lanes:

Western blot - 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) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 23 kDa

Observed band size: 14 kDa,17 kDa

true

Exposure time: 60s

• WB

Lab

Western blot - Anti-Frataxin antibody [EPR21840] (AB219414)

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

The bands detected by this product are consistent with publications PMID : 18725397 and PMID : 20156111.

We suggest optimizing experimental protocols (increasing lysate amount, using lower dilution or higher sensitivity ECL substrate) to improve results when using ab197896 in western blot.

All lanes:

Western blot - Anti-Frataxin antibody [EPR21840] (ab219414) at 1/1000 dilution

Lane 1:

Human liver tissue lysate at 20 µg

Lane 2:

Mouse liver tissue lysate at 20 µg

Lane 3:

SW480 (human colorectal adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 4:

TF-1 (human Erythroleukemia erythroblast) whole cell lysate at 20 µg

Lane 5:

Huh7 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 6:

HCT116 (human colorectal carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 7:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 8:

RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 9:

PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 14 kDa,16 kDa,19 kDa

false

• WB

Lab

Western blot - Anti-Frataxin antibody [EPR21840] (AB219414)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. ab213204 was used as laoding control.

All lanes:

Western blot - 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) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 23 kDa

Observed band size: 16 kDa

false

Exposure time: 3s