Anti-GALT antibody [EPR12555]

• IP

Supplier Data

Immunoprecipitation - Anti-GALT antibody [EPR12555] (AB178406)

Western blot analysis on Immunoprecipitation pellet from either 1) Human fetal liver lysate, or 2) 1xPBS (negative control); showing GALT, immunoprecipitated using ab178406 at 1/10 dilution with HRP-conjugated anti-rabbit IgG preferentially detecting the non-reduced form of rabbit IgG.

All lanes:

Immunoprecipitation - Anti-GALT antibody [EPR12555] (ab178406)

Predicted band size: 43 kDa

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• WB

Lab

Western blot - Anti-GALT antibody [EPR12555] (AB178406)

Lanes 1-4 : Merged signal (red and green). Green - ab178406 observed at 43 kDa. Red - loading control ab7291 observed at 50 kDa.

ab178406 Anti-GALT antibody [EPR12555] was shown to specifically react with Galactose-1-Phosphate Uridylyltransferase in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265027 (knockout cell lysate ab257958) was used. Wild-type and Galactose-1-Phosphate Uridylyltransferase knockout samples were subjected to SDS-PAGE. ab178406 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) 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-GALT antibody [EPR12555] (ab178406) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

GALT knockout HeLa cell lysate at 20 µg

Lane 3:

HepG2 cell lysate at 20 µg

Lane 4:

A549 cell lysate at 20 µg

Predicted band size: 43 kDa

Observed band size: 43 kDa

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• WB

Lab

Western blot - Anti-GALT antibody [EPR12555] (AB178406)

Lanes 1 and 2 : Green signal from target – ab178406 observed at 43 kDa

Lanes 3 and 4 : Red signal from loading control – ab8245 observed at 37 kDa

Lanes 5 and 6 : Merged (red and green) signal

ab178406 was shown to react with GALT when GALT knockout samples were used, along with additional cross-reactive bands. Wild-type and GALT knockout samples were subjected to SDS-PAGE. ab178406 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/2000 respectively and incubated overnight at 4°C. Blots were developed withGoat anti-Rabbit IgG H&L (IRDye® 800CW)preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776)secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.

Lanes 1 - 2:

Western blot - Anti-GALT antibody [EPR12555] (ab178406) at 1/1000 dilution

Lanes 3 - 4:

Western blot - Anti-GALT antibody [EPR12555] (ab178406) at 1/2000 dilution

Lanes 1 and 3:

GALT knockout HAP1 cell lysate at 20 µg

Lanes 2 and 4:

Wild-type HAP1 cell lysate at 20 µg

Predicted band size: 43 kDa

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• WB

Supplier Data

Western blot - Anti-GALT antibody [EPR12555] (AB178406)

All lanes:

Western blot - Anti-GALT antibody [EPR12555] (ab178406) at 1/1000 dilution

Lane 1:

K562 cell lysate at 10 µg

Lane 2:

MDA-MB-435 cell lysate at 10 µg

Lane 3:

Human fetal brain lysate at 10 µg

Lane 4:

Human fetal liver lysate at 10 µg

Lane 5:

HepG2 cell lysate at 10 µg

Secondary

All lanes:

Goat anti-rabbit HRP at 1/2000 dilution

Predicted band size: 43 kDa

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