Anti-ETFDH antibody [3D1AC4AF3]

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ETFDH antibody [3D1AC4AF3] (AB131376)

IHC image of ETFDH staining in Human normal heart muscle formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. 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 ab131376, 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.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-ETFDH antibody [3D1AC4AF3] (AB131376)

Immunofluorescent analysis of ETFDH in HDFn cells.
The cells were paraformaldehyde fixed (4%, 20 minutes) and Triton X-100 permeabilized (0.1%, 15 minutes). The cells were then incubated with ab131376 at 5 µg/ml for 2 h at room temperature, or over night at 4°C. The secondary antibody was (red) Alexa Fluor® 594 Goat anti-Mouse IgG (H+L) used at a 1/1000 dilution for 1 h. 1% BSA was used as the blocking agent for all blocking steps. The target protein locates to the mitochondria.

• Flow Cyt

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Flow Cytometry - Anti-ETFDH antibody [3D1AC4AF3] (AB131376)

Flow cytometric analysis of ETFDH in HeLa cells fixed and permeabilized with methanol and stained with 1 µg/mL of ab131376 (blue), or an equal amount of an isotype control antibody (red). 1% BSA was used as the blocking reagent for all the blocking steps.

• WB

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Western blot - Anti-ETFDH antibody [3D1AC4AF3] (AB131376)

The high background signal in Mouse tissue sample was caused by the direct reaction between the Mouse IgG in Mouse tissue preps and the Goat anti-Mouse secondary antibody.

All lanes:

Western blot - Anti-ETFDH antibody [3D1AC4AF3] (ab131376) at 1 µg/mL

Lane 1:

Molecular weight marker

Lane 2:

Human Liver Homogenate (HLH) at 10 µg

Lane 3:

Human Heart Homogenate (HHH) at 10 µg

Lane 4:

HepG2 Cell Lysate at 15 µg

Lane 5:

Hela Cell Lysate at 15 µg

Lane 6:

HDFn Cell Lysate at 15 µg

Lane 7:

SY5Y Cell Lysate at 15 µg

Lane 8:

Jurkat Cell Lysate at 15 µg

Lane 9:

Mouse Liver Homogenate (MLH) at 10 µg

Lane 10:

Mouse Heart Homogenate (MHH) at 10 µg

Lane 11:

Rat Liver Homogenate (RLH) at 10 µg

Lane 12:

Rat Heart Homogenate (RHH) at 10 µg

Lane 13:

Rat Brain Homogenate (RBH) at 10 µg

Lane 14:

Rat Kidney Homogenate (RLH) at 10 µg

Lane 15:

Rat Skeletal Muscle Homogenate (RSMH) at 10 µg

Secondary

All lanes:

Goat anti-Mouse AP at 1/3000 dilution

Predicted band size: 68 kDa

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