Anti-BCKDK antibody [EPR7564]

• WB

Lab

Western blot - Anti-BCKDK antibody [EPR7564] (AB128935)

Lane 1 : Wild-type HAP1 cell lysate (20 μg)
Lane 2 : BCKDH kinase knockout HAP1 cell lysate (20 μg)
Lane 3 : HeLa cell lysate (20 μg)
Lanes 1 - 3 : Merged signal (red and green). Green - ab128935 observed at 48 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab128935 was shown to specifically react with BCKDH kinase when BCKDH kinase knockout samples were used. Wild-type and BCKDH kinase knockout samples were subjected to SDS-PAGE. ab128935 and ab8245 (loading control to GAPDH) were diluted at 1/1000 and 1/10,000 dilution respectively and incubated overnight at 4C. 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 1/10,000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-BCKDK antibody [EPR7564] (ab128935)

Predicted band size: 46 kDa

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• WB

Lab

Western blot - Anti-BCKDK antibody [EPR7564] (AB128935)

All lanes:

Western blot - Anti-BCKDK antibody [EPR7564] (ab128935) at 1/1000 dilution

Lane 1:

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

Lane 2:

Rat heart tissue lysate at 20 µg

Lane 3:

Rat kidney tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 46 kDa

Observed band size: 46 kDa

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• WB

Lab

Western blot - Anti-BCKDK antibody [EPR7564] (AB128935)

All lanes:

Western blot - Anti-BCKDK antibody [EPR7564] (ab128935) at 1/1000 dilution

Lane 1:

PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate at 15 µg

Lane 2:

RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 46 kDa

Observed band size: 46 kDa

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• WB

Unknown

Western blot - Anti-BCKDK antibody [EPR7564] (AB128935)

All lanes:

Western blot - Anti-BCKDK antibody [EPR7564] (ab128935) at 1/1000 dilution

Lane 1:

Fetal kidney lysate at 10 µg

Lane 2:

HeLa cell lysate at 10 µg

Lane 3:

HepG2 cell lysate at 10 µg

Lane 4:

SH-SY5Y cell lysate at 10 µg

Secondary

All lanes:

Standard HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 46 kDa

Observed band size: 46 kDa

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• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-BCKDK antibody [EPR7564] (AB128935)

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.