Anti-Mitofusin 2 antibody [NIAR164] - BSA and Azide free

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mitofusin 2 antibody [NIAR164] - BSA and Azide free (AB219730)

Unpurified ab124773, at 1/50, staining Mitofusin 2 in formalin fixed paraffin embedded Human kidney tissue using immunohistochemistry.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab124773).

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Mitofusin 2 antibody [NIAR164] - BSA and Azide free (AB219730)

Immunofluorescence staining of HEK293 cells with purified ab124773 at a working dilution of 1/300, counter-stained with DAPI. The secondary antibody was Alexa Fluor^® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor^® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 100% methanol and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab124773 was used at a dilution of 1/500 followed by an Alexa Fluor^® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab124773).

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mitofusin 2 antibody [NIAR164] - BSA and Azide free (AB219730)

Immunohistochemical staining of paraffin embedded human kidney with purified ab124773 at a working dilution of 1/300. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab124773).

• WB

Lab

Western blot - Anti-Mitofusin 2 antibody [NIAR164] - BSA and Azide free (AB219730)

This data was developed using ab124773, the same antibody clone in a different buffer formulation.

Western blot : Anti-MFN2 antibody [NIAR164] (ab124773) staining at 1/5000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab124773 was shown to bind specifically to MFN2. A band was observed at 64 kDa in wild-type HEK-293 cell lysates with no signal observed at this size in MFN2 knockout cell line. To generate this image, wild-type and MFN2 knockout HEK-293 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-Mitofusin 2 antibody [NIAR164] (<a href='/en-us/products/primary-antibodies/mitofusin-2-antibody-niar164-ab124773'>ab124773</a>) at 1/5000 dilution

Lane 1:

Wild-type HEK-293 cell lysate at 20 µg

Lane 2:

Western blot - Human MFN2 (Mitofusin 2) knockout HEK-293 cell line (<a href='/en-us/products/cell-lines/human-mfn2-mitofusin-2-knockout-hek-293-cell-line-ab260861'>ab260861</a>)

Lane 2:

MFN2 knockout HEK-293 cell lysate at 20 µg

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

PC-3 cell lysate at 20 µg

Secondary

Lanes 1 - 4:

Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution

Lanes 1 - 4:

Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 64 kDa

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