Anti-Glycerol kinase antibody [EPR6567]

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Glycerol kinase antibody [EPR6567] (AB126599)

ab126599, at 1/100 dilution, staining Glycerol kinase in permeabilized HepG2 cells by ImmunoFluorescence (red). A rabbit IgG is used as a negative control (green).

• WB

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Western blot - Anti-Glycerol kinase antibody [EPR6567] (AB126599)

All lanes:

Western blot - Anti-Glycerol kinase antibody [EPR6567] (ab126599) at 1/1000 dilution

Lane 1:

Fetal liver tissue lysate at 10 µg

Lane 2:

HepG2 cell lysate at 10 µg

Secondary

All lanes:

Goat anti-rabbit HRP at 1/2000 dilution

Predicted band size: 61 kDa

Observed band size: 55 kDa

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• WB

Lab

Western blot - Anti-Glycerol kinase antibody [EPR6567] (AB126599)

Lanes 1-4 : Merged signal (red and green). Green - ab126599 observed at 61 kDa. Red - loading control ab8245 observed at 36 kDa.

ab126599 Anti-Glycerol kinase antibody [EPR6567] was shown to specifically react with Glycerol kinase in wild-type HEK-293T cells. Loss of signal was observed when knockout cell line ab267328 (knockout cell lysate ab257966) was used. Wild-type and Glycerol kinase knockout samples were subjected to SDS-PAGE. ab126599 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 500 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-Glycerol kinase antibody [EPR6567] (ab126599) at 1/500 dilution

Lane 1:

Wild-type HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg

Lane 2:

GK knockout HEK-293T (Human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 20 µg

Lane 2:

Western blot - Human GK (Glycerol kinase) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-gk-glycerol-kinase-knockout-hek-293t-cell-line-ab267328'>ab267328</a>)

Lane 3:

HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg

Lane 4:

Jurkat (Human T cell leukemia cell line from peripheral blood) whole 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: 61 kDa

Observed band size: 61 kDa

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• WB

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Western blot - Anti-Glycerol kinase antibody [EPR6567] (AB126599)

All lanes:

Western blot - Anti-Glycerol kinase antibody [EPR6567] (ab126599) at 1/1000 dilution

All lanes:

Mouse adipose tissue lysate at 25 µg

Secondary

All lanes:

HRP-conjugated Donkey anti-rabbit polyclonal at 1/5000 dilution

Predicted band size: 61 kDa

Observed band size: 55 kDa

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Exposure time: 5min

This image is courtesy of an anonymous Abreview

• OI-RD Scanning

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OI-RD Scanning - Anti-Glycerol kinase antibody [EPR6567] (AB126599)

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.