Anti-Mitofusin 2 antibody [NIAR164]

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Mitofusin 2 antibody [NIAR164] (AB124773)

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.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mitofusin 2 antibody [NIAR164] (AB124773)

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

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

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Mitofusin 2 antibody [NIAR164] (AB124773)

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.

• WB

Lab

Western blot - Anti-Mitofusin 2 antibody [NIAR164] (AB124773)

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] (ab124773) 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

false

• WB

Lab

Western blot - Anti-Mitofusin 2 antibody [NIAR164] (AB124773)

Blocking buffer : 5% NFDM/TBST
Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-Mitofusin 2 antibody [NIAR164] (ab124773) at 1/5000 dilution

Lane 1:

HeLa cell lysate at 20 µg

Lane 2:

Jurkat cell lysate at 20 µg

Lane 3:

HEK293 cell lysate at 20 µg

Lane 4:

Raji cell lysate at 20 µg

Secondary

All lanes:

HRP goat anti-rabbit IgG (H+L) at 1/50000 dilution

Predicted band size: 124 kDa,86 kDa

Observed band size: 124 kDa,80 kDa

false

• WB

Lab

Western blot - Anti-Mitofusin 2 antibody [NIAR164] (AB124773)

Blocking buffer : 5% NFDM/TBST
Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-Mitofusin 2 antibody [NIAR164] (ab124773) at 1/5000 dilution

Lane 1:

mouse brain lysate at 20 µg

Lane 2:

mouse kidney lysate at 20 µg

Lane 3:

rat brain lysate at 20 µg

Secondary

All lanes:

HRP goat anti-rabbit IgG (H+L) at 1/50000 dilution

Predicted band size: 86 kDa

Observed band size: 80 kDa

false

• WB

AbReview36157****

Western blot - Anti-Mitofusin 2 antibody [NIAR164] (AB124773)

All lanes:

Western blot - Anti-Mitofusin 2 antibody [NIAR164] (ab124773) at 1/1000 dilution

All lanes:

Rat primary neurons cell lysate at 20 µg

Secondary

All lanes:

Anti-rabbit IgG HRP conjugate at 1/2000 dilution

Predicted band size: 86 kDa

true

Exposure time: 30s

This image is courtesy of an anonymous Abreview