Anti-COMT antibody [EPR6491(B)]

• Flow Cyt

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Flow Cytometry - Anti-COMT antibody [EPR6491(B)] (AB124813)

ab124813, at 1/10 dilution staining COMT in A431 cells, (Red) compared to a nonspecific negative control antibody (Green) by Flow Cytometry.

• WB

Lab

Western blot - Anti-COMT antibody [EPR6491(B)] (AB124813)

Lanes 1 - 4 : Merged signal (red and green). Green - ab124813 observed at 28 kDa. Red - loading control, ab18058, observed at 130 kDa.

ab124813 was shown to recognize COMT when COMT knockout samples were used, along with additional cross-reactive bands. Wild-type and COMT knockout samples were subjected to SDS-PAGE. ab124813 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10000 dilution respectively. Blots were developed with 800CW Goat anti Rabbit and 680CW Goat anti Mouse secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-COMT antibody [EPR6491(B)] (ab124813) at 1/1000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

COMT knockout HAP1 whole cell lysate at 20 µg

Lane 3:

A431 whole cell lysate at 20 µg

Lane 4:

MOLT4 whole cell lysate at 20 µg

Predicted band size: 30 kDa

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• WB

Lab

Western blot - Anti-COMT antibody [EPR6491(B)] (AB124813)

Lanes 1-2 : Merged signal (red and green). Green - ab124813 observed at 28 kDa. Red - loading control ab8245 observed at 37 kDa.

ab124813 Anti-COMT antibody [EPR6491(B)] was shown to specifically react with COMT in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab266537 (knockout cell lysate ab257396) was used. Wild-type and COMT knockout samples were subjected to SDS-PAGE. ab124813 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-COMT antibody [EPR6491(B)] (ab124813) at 1/1000 dilution

Lane 1:

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

Lane 2:

COMT knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human COMT knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-comt-knockout-hek-293t-cell-line-ab266537'>ab266537</a>)

Predicted band size: 30 kDa

Observed band size: 28 kDa

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• WB

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Western blot - Anti-COMT antibody [EPR6491(B)] (AB124813)

All lanes:

Western blot - Anti-COMT antibody [EPR6491(B)] (ab124813) at 1/1000 dilution

Lane 1:

Human fetal liver lysate at 10 µg

Lane 2:

A431 cell lysate at 10 µg

Lane 3:

MOLT4 cell lysate at 10 µg

Predicted band size: 30 kDa

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• OI-RD Scanning

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OI-RD Scanning - Anti-COMT antibody [EPR6491(B)] (AB124813)

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.