Anti-Glucose 6 Phosphate Dehydrogenase antibody

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Glucose 6 Phosphate Dehydrogenase antibody (AB993)

ICC/IF image of ab993 stained MCF7 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab993 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

• IP

Supplier Data

Immunoprecipitation - Anti-Glucose 6 Phosphate Dehydrogenase antibody (AB993)

Glucose 6 Phosphate Dehydrogenase was immunoprecipitated from NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate (1 mg for IP, 20% of IP loaded) with ab993 at 6 μg/mg lysate. Western blot was performed from the immunoprecipitate using ab993 at 1 μg/ml.

Lane 1 : ab993 (batch 3) IP in NIH/3T3 whole cell lysate.

Lane 2 : ab993 (batch 4) IP in NIH/3T3 whole cell lysate.

Lane 3 : Control IgG IP in NIH/3T3 whole cell lysate.

Detection : Chemiluminescence with exposure time of 30 seconds.

All lanes:

Immunoprecipitation - Anti-Glucose 6 Phosphate Dehydrogenase antibody (ab993)

Predicted band size: 59 kDa

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• WB

Supplier Data

Western blot - Anti-Glucose 6 Phosphate Dehydrogenase antibody (AB993)

All lanes:

Western blot - Anti-Glucose 6 Phosphate Dehydrogenase antibody (ab993) at 1 µg/mL

Lane 1:

HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 50 µg

Lane 2:

HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) whole cell lysate at 50 µg

Lane 3:

NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate at 50 µg

Predicted band size: 59 kDa

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Exposure time: 30s

• WB

CiteAb

Western blot - Anti-Glucose 6 Phosphate Dehydrogenase antibody (AB993)

Western Blotting using Anti-Glucose 6 Phosphate Dehydrogenase antibody, ab993. Publication image from Serrano, M. et al., 2016, Nat Commun, 26976705. Legend direct from paper.

Characterization of the G6PD-Tg mice.(a) Relative G6PD mRNA of the indicated tissues from mice 10–12 weeks old, one male and two females for the WT group, one male (M) and five females (F) in the Tg group. (b) Levels of G6PD protein in the indicated tissues from 3 to 4-months-old mice and mouse embryo fibroblasts (MEFs) where each lane corresponds to one individual animal for the tissues (WAT : white adipose tissue; RBC : red blood cells), or an independent MEF preparation for the MEFs. The anti-G6PD antibody reacts with both the mouse and human G6PD protein. In some tissues (liver, heart), a second upper band was also detected. ACTIN or GAPDH were used as loading controls. (c) G6PD enzymatic activity in the indicated samples. (d) Levels of NADPH in the indicated tissues. (e) Uric acid concentration in plasma. (f) Lactate production in isolated erythrocytes (left panel) and total levels of lactate in plasma (right panel). (g,h) Percentage of cell survival in G6PD-Tg and WT primary MEFs upon treatment with the oxidative stress inducers diamide (g) and paraquat (h). (i) Survival curve of mice treated with a lethal dose of paraquat. Bars represent the mean±s.e.m. Statistic significance was assessed using the two-tailed Student's t-test for all experiments, with the exception of i, where the logrank test was applied : *P<0.05; **P<0.01; ***P<0.001.

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