Anti-Lipoamide Dehydrogenase antibody [EPR6634(B)]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lipoamide Dehydrogenase antibody [EPR6634(B)] (AB124926)

ab124926 at 1/100 dilution staining Lipoamide Dehydrogenase in Paraffin-embedded Human colon tissue by Immunohistochemistry.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• WB

Lab

Western blot - Anti-Lipoamide Dehydrogenase antibody [EPR6634(B)] (AB124926)

Lanes 1-3 : Merged signal (red and green). Green - ab124926 observed at 56 kDa. Red - loading control ab8245 observed at 36 kDa.

ab124926 Anti-Lipoamide Dehydrogenase antibody [EPR6634(B)] was shown to specifically react with Lipoamide Dehydrogenase in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265535 (knockout cell lysate ab257922) was used. Wild-type and Lipoamide Dehydrogenase knockout samples were subjected to SDS-PAGE. ab124926 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-Lipoamide Dehydrogenase antibody [EPR6634(B)] (ab124926) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

DLD knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human DLD (Lipoamide Dehydrogenase) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-dld-lipoamide-dehydrogenase-knockout-hela-cell-line-ab265535'>ab265535</a>)

Lane 3:

Jurkat cell 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: 54 kDa

Observed band size: 56 kDa

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• WB

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Western blot - Anti-Lipoamide Dehydrogenase antibody [EPR6634(B)] (AB124926)

All lanes:

Western blot - Anti-Lipoamide Dehydrogenase antibody [EPR6634(B)] (ab124926) at 1/1000 dilution

Lane 1:

Jurkat cell lysate at 10 µg

Lane 2:

MCF7 cell lysate at 10 µg

Lane 3:

HeLa cell lysate at 10 µg

Lane 4:

293T cell lysate at 10 µg

Secondary

All lanes:

HRP-conjugated goat anti-rabbit at 1/2000 dilution

Predicted band size: 54 kDa

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• OI-RD Scanning

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OI-RD Scanning - Anti-Lipoamide Dehydrogenase antibody [EPR6634(B)] (AB124926)

We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody. Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.