Anti-Lactate Dehydrogenase B/LDH-B antibody [EP1565Y]

• WB

Lab

Western blot - Anti-Lactate Dehydrogenase B/LDH-B antibody [EP1565Y] (AB53292)

Lanes 1- 2 : Merged signal (red and green). Green - ab53292 observed at 37 kDa. Red - Anti-Vinculin antibody [VIN-54] observed at 124 kDa.

ab53292 was shown to react with Lactate Dehydrogenase B/LDH-B in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab255403 (knockout cell lysate ab263761) was used. Wild-type HEK-293T and LDHB knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab53292 and Anti-Vinculin antibody [VIN-54] overnight at 4°C at a 1 in 5000 dilution and a 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-Lactate Dehydrogenase B/LDH-B antibody [EP1565Y] (ab53292) at 1/5000 dilution

Lane 1:

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

Lane 2:

LDHB knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human LDHB (Lactate Dehydrogenase B) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-ldhb-lactate-dehydrogenase-b-knockout-hek-293t-cell-line-ab255403'>ab255403</a>)

Predicted band size: 37 kDa

Observed band size: 37 kDa

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• WB

Lab

Western blot - Anti-Lactate Dehydrogenase B/LDH-B antibody [EP1565Y] (AB53292)

Lanes 1 - 3 : Merged signal (red and green). Green - ab53292 observed at 37 kDa. Red - loading control, ab130007, observed at 130 kDa.

ab53292 was shown to specifically react with Lactate Dehydrogenase B in wild-type HAP1 cells as signal was lost in LDHB (Lactate Dehydrogenase B) knockout cells. Wild-type and LDHB (Lactate Dehydrogenase B) knockout samples were subjected to SDS-PAGE. ab53292 and ab130007 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1/50000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Lactate Dehydrogenase B/LDH-B antibody [EP1565Y] (ab53292) at 1/50000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

LDHB (Lactate Dehydrogenase B) knockout HAP1 whole cell lysate at 20 µg

Lane 3:

HeLa whole cell lysate at 20 µg

Predicted band size: 37 kDa

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• WB

Unknown

Western blot - Anti-Lactate Dehydrogenase B/LDH-B antibody [EP1565Y] (AB53292)

All lanes:

Western blot - Anti-Lactate Dehydrogenase B/LDH-B antibody [EP1565Y] (ab53292)

All lanes:

HeLa cell lysate at 10 µg

Secondary

All lanes:

Goat anti-rabbit HRP

Predicted band size: 37 kDa

Observed band size: 37 kDa

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