Anti-CBS antibody [EPR8578]

• WB

Lab

Western blot - Anti-CBS antibody [EPR8578] (AB131155)

Lanes 1-3 : Merged signal (red and green). Green - ab131155 observed at 70 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab131155 Anti-CBS antibody [EPR8578] was shown to specifically react with CBS in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264950 (knockout cell lysate ab257203) was used. Wild-type and CBS knockout samples were subjected to SDS-PAGE. ab131155 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 10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-CBS antibody [EPR8578] (ab131155) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

CBS knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human CBS knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-cbs-knockout-hela-cell-line-ab264950'>ab264950</a>)

Lane 3:

Raji cell lysate at 20 µg

Predicted band size: 61 kDa

Observed band size: 70 kDa

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• WB

Unknown

Western blot - Anti-CBS antibody [EPR8578] (AB131155)

Lanes 1-3 : Merged signal (red and green). Green - ab131155 observed at 70 kDa. Red - loading control, ab8245 observed at 37 kDa.

ab131155 Anti-CBS antibody [EPR8578] was shown to specifically react with CBS in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264950 (knockout cell lysate ab257203) was used. Wild-type and CBS knockout samples were subjected to SDS-PAGE. ab131155 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 10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-CBS antibody [EPR8578] (ab131155) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

CBS knockout HeLa cell lysate at 20 µg

Lane 3:

Raji cell lysate at 20 µg

Predicted band size: 61 kDa

Observed band size: 70 kDa

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• WB

Lab

Western blot - Anti-CBS antibody [EPR8578] (AB131155)

Blocking and diluting buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-CBS antibody [EPR8578] (ab131155) at 1/1000 dilution

Lane 1:

Human liver tissue lysate at 20 µg

Lane 2:

Raji (Human Burkitt's lymphoma B lymphocyte) whole cell lysate at 20 µg

Lane 3:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

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Exposure time: 20s

• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-CBS antibody [EPR8578] (AB131155)

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.