Anti-SOD2/MnSOD antibody [EPR2560Y]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SOD2/MnSOD antibody [EPR2560Y] (AB68155)

Immunohistochemical analysis of paraffin-embedded human cardiac muscle tissue labeling SOD2/MnSOD with ab68155 at 1/4000 dilution.

Positive staining on human cardiac muscle.

The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument

Secondary was a ready-to-use LeicaDS9800 (Bond™ Polymer Refine Detection) kit.

Counterstained with Hematoxylin.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SOD2/MnSOD antibody [EPR2560Y] (AB68155)

Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling SOD2/MnSOD with ab68155 at 1/1000 dilution.

Positive staining on rat kidney.

The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument

Secondary was a ready-to-use LeicaDS9800 (Bond™ Polymer Refine Detection) kit.

Counterstained with Hematoxylin.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-SOD2/MnSOD antibody [EPR2560Y] (AB68155)

Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling SOD2/MnSOD with ab68155 at 1/1000 dilution.

Positive staining on mouse kidney.

The primary antibody was incubated for 30 mins at room temperature.

The immunostaining was performed on a Leica Biosystems BOND^® RX instrument

Secondary was a ready-to-use LeicaDS9800 (Bond™ Polymer Refine Detection) kit.

Counterstained with Hematoxylin.

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• WB

Unknown

Western blot - Anti-SOD2/MnSOD antibody [EPR2560Y] (AB68155)

Lanes 1 - 4 : Merged signal (red and green). Green - ab68155 observed at 29 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab68155 was shown to specifically react with SOD2 when SOD2 knockout samples were used. Wild-type and SOD2 knockout samples were subjected to SDS-PAGE. ab68155 and ab8245 (loading control to GAPDH) were diluted at 1/1000 and 1/10 000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-SOD2/MnSOD antibody [EPR2560Y] (ab68155) at 1/1000 dilution

Lane 1:

Wild-type HAP1 cell lysate at 20 µg

Lane 2:

SOD2 knockout HAP1 cell lysate at 20 µg

Lane 3:

Human heart tissue lysate at 20 µg

Lane 4:

Rat heart tissue lysate at 20 µg

Predicted band size: 24 kDa

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• WB

Unknown

Western blot - Anti-SOD2/MnSOD antibody [EPR2560Y] (AB68155)

All lanes:

Western blot - Anti-SOD2/MnSOD antibody [EPR2560Y] (ab68155) at 1/500 dilution

Lane 1:

Rat brain lysate at 10 µg

Lane 2:

SH-SY5Y lysate at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 24 kDa

Observed band size: 25 kDa

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• WB

Lab

Western blot - Anti-SOD2/MnSOD antibody [EPR2560Y] (AB68155)

Lanes 1 - 4 : Merged signal (red and green). Green - ab68155 observed at 22 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab68155 was shown to react with SOD2/MnSOD in wild-type HEK-293T. Loss of signal was observed when knockout cell line ab255434 (knockout cell lysate ab263835) was used. Wild-type and SOD2/MnSOD knockout samples were subjected to SDS-PAGE. ab68155 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-SOD2/MnSOD antibody [EPR2560Y] (ab68155) at 1/1000 dilution

Lane 1:

SHSY5Y cell lysate at 20 µg

Lane 2:

A549 cell lysate at 20 µg

Lane 3:

Wild-type HEK-293T cell lysate at 20 µg

Lane 4:

Western blot - Human SOD2 (MnSOD) knockout HEK-293T cell lysate (<a href='/ko/products/cell-lysates/human-sod2-mnsod-knockout-hek-293t-cell-lysate-ab263835'>ab263835</a>) at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution

Predicted band size: 24 kDa

Observed band size: 29 kDa,37 kDa

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• WB

Lab

Western blot - Anti-SOD2/MnSOD antibody [EPR2560Y] (AB68155)

This blot was produced using 4-20% SDS-PAGE containing 15 μg of rat brain lysate per lane at 150V for 1hr before being transferred onto a 0.45 μm PVDF membrane at 75V for 1hr. The membrane was then blocked for 1hr using 5% NFDM/TBST, then incubated with ab68155 (1/1000) at room temperature for 1hr. After being washed three times in TBST, the membrane was incubated with Peroxidase conjugated goat anti-rabbit IgG (H+L) (ab97051) at 1/100,000 dilution for 1hr at room temperature. The membrane was washed three times again. Then the signal was developed using the ECL technique.

ab68155 was stored at a range of temperatures (+4°C, +22°C, +37°C) for 1 week before being tested in WB. The image shows the band intensity remains relatively constant across all storage temperatures, demonstrating that antibody activity is not affected under these conditions. This data was generated as part of a structured antibody stability study supporting Abcam’s transition from cold‑chain to ambient shipping.

All lanes:

Western blot - Anti-SOD2/MnSOD antibody [EPR2560Y] (ab68155) at 1/1000 dilution

All lanes:

Rat brain lysate at 15 µg with NDFM/TBST

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

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Exposure time: 7s