Anti-MDH2 antibody [EPR14883(B)]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-MDH2 antibody [EPR14883(B)] (AB181857)

Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed HeLa cells labeling MDH2 with ab181857 at 1/10 dilution (red) compared to a Rabbit IgG monoclonal isotype control (green), followed by Goat anti rabbit IgG (FITC) secondary antibody at 1/150 dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MDH2 antibody [EPR14883(B)] (AB181857)

Immunohistochemical analysis of paraffin-embedded Human hepatocellular carcinoma tissue labeling MDH2 with ab181857 at 1/50 dilution followed by prediluted HRP Polymer for Rabbit IgG. Counter stained with Hematoxylin.

Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

• WB

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Western blot - Anti-MDH2 antibody [EPR14883(B)] (AB181857)

All lanes:

Western blot - Anti-MDH2 antibody [EPR14883(B)] (ab181857) at 1/50000 dilution

Lane 1:

K562 cell lysate at 20 µg

Lane 2:

HepG2 cell lysate at 20 µg

Lane 3:

HeLa cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution

Predicted band size: 36 kDa

Observed band size: 36 kDa

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• WB

Lab

Western blot - Anti-MDH2 antibody [EPR14883(B)] (AB181857)

Lanes 1-4 : Merged signal (red and green). Green - ab181857 observed at 36 kDa. Red - loading control ab7291 observed at 50 kDa.

ab181857 Anti-MDH2 antibody [EPR14883(B)] was shown to specifically react with MDH2 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266449 (knockout cell lysate ab257533) was used. Wild-type and MDH2 knockout samples were subjected to SDS-PAGE. ab181857 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) 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-MDH2 antibody [EPR14883(B)] (ab181857) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

MDH2 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human MDH2 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-mdh2-knockout-hek-293t-cell-line-ab266449'>ab266449</a>)

Lane 3:

K-562 cell lysate at 20 µg

Lane 4:

Human eyeball tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 36 kDa

Observed band size: 36 kDa

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