Anti-MPP1 antibody [EPR5864]

• WB

Lab

Western blot - Anti-MPP1 antibody [EPR5864] (AB133500)

Lanes 1-4 : Merged signal (red and green). Green - ab133500 observed at 52 kDa. Red - loading control ab8245 observed at 36 kDa.

ab133500 Anti-MPP1 antibody [EPR5864] was shown to specifically react with MPP1 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab267248 (knockout cell lysate ab258051) was used. Wild-type and MPP1 knockout samples were subjected to SDS-PAGE. ab133500 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-MPP1 antibody [EPR5864] (ab133500) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

MPP1 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human MPP1 knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-mpp1-knockout-hek-293t-cell-line-ab267248'>ab267248</a>)

Lane 3:

Jurkat cell lysate at 20 µg

Lane 4:

K-562 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: 52 kDa

Observed band size: 52 kDa

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• WB

Unknown

Western blot - Anti-MPP1 antibody [EPR5864] (AB133500)

All lanes:

Western blot - Anti-MPP1 antibody [EPR5864] (ab133500) at 1/1000 dilution

Lane 1:

HeLa cell lysate at 10 µg

Lane 2:

293T cell lysate at 10 µg

Lane 3:

Jurkat cell lysate at 10 µg

Lane 4:

Human heart lysate at 10 µg

Lane 5:

K562 cell lysate at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 52 kDa

Observed band size: 55 kDa

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• OI-RD Scanning

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OI-RD Scanning - Anti-MPP1 antibody [EPR5864] (AB133500)

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.