Anti-PYGL antibody - C-terminal

• IP

Supplier Data

Immunoprecipitation - Anti-PYGL antibody - C-terminal (AB190243)

Immunoprecipitation analysis of 293T cell lysate (in RIPA buffer, 0.5 or 1.0 mg per IP reaction; 20% of IP loaded) labeling PYGL using ab190243 at 6 μg per reaction (lane 1). a Control IgG was used in lane 2. For blotting immunoprecipitated PYGL, ab190243 was used at 1 μg/ml. Detection : Chemiluminescence with an exposure time of 30 seconds.

All lanes:

Immunoprecipitation - Anti-PYGL antibody - C-terminal (ab190243)

Predicted band size: 97 kDa

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• WB

Supplier Data

Western blot - Anti-PYGL antibody - C-terminal (AB190243)

All lanes:

Western blot - Anti-PYGL antibody - C-terminal (ab190243) at 0.1 µg/mL

Lane 1:

HeLa whole cell lysate at 50 µg

Lane 2:

293T whole cell lysate at 50 µg

Lane 3:

Jurkat whole cell lysate at 50 µg

Lane 4:

TCMK-1 whole cell lysate at 50 µg

Lane 5:

NIH 3T3 whole cell lysate at 50 µg

Predicted band size: 97 kDa

true

Exposure time: 3min

• WB

CiteAb

Western blot - Anti-PYGL antibody - C-terminal (AB190243)

PYGL western blot using anti-PYGL antibody - C-terminal ab190243. Publication image and figure legend from Ma, J., Wei, K., et al., 2020, Nat Commun, PubMed 32286295.

ab190243 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab190243 please see the product overview.

Glycogenolysis-derived G6P is channeled to the PPP.a, b Pygl and Pygm expression in untreated, IFN-γ/LPS or IL-4 treated BMDMs were determined by real-time PCR (a) and western blot (b). c BMDMs differentiated in normal 12C-glucose were stimulated with IFN-γ/LPS or IL-4 for 6 h and switched to 13C-glucose for 6 h, LC-MS/MS was performed for m + 5-labeled R5P. d BMDMs were pretreated with GPI for 30 min or pretransfected with siRNA (Gys1 or Pygl) for 24 h prior to stimulation with IFN-γ/LPS for 6 h and switched to 13C-glucose for 6 h, LC-MS/MS was performed for m + 5-labeled R5P. e, f G6pdx and 6Pgd expression in untreated, IFN-γ/LPS or IL-4 treated BMDMs were determined by real-time PCR (e) and western blot (f). gPygl or G6pdx siRNA transfected BMDMs were stimulated with IFN-γ/LPS for 36 h. Intracellular glycogen levels were detected by colorimetric assay. h BMDMs differentiated in normal 12C-glucose were stimulated with IFN-γ/LPS or IL-4 for 6 h and switched to 13C-glucose for 6 h, LC-MS/MS was performed for m + 7-labeled S7P, m + 5-labeled S7P and m + 4-labeled E4P. i, j BMDMs cultured and differentiated in 13C-glucose were pretreated with GPI for 30 min prior to stimulation with IFN-γ/LPS for 6 h and switched to 12C-glucose for 2 or 4 h, 13C-labeled G6P/G1P were detected by LC-MS/MS (i). The ratio of G6P/G1P from glycogenolysis or glycolysis was calculated. The glycogenolysis-derived G6P using the format of [m + 6 G6P (IFN-γ/LPS)–m + 6 G6P (IFN-γ/LPS + GPI)]/Total G6P (IFN-γ/LPS) and the glycolysis-derived G6P by the format of [m + 0 G6P (IFN-γ/LPS)] / Total G6P (IFN-γ/LPS) (j). Unless otherwise specified, n = 3 biologically independent experiments were performed. Data are presented as mean ± SEM. P values were calculated using one-way ANOVA.

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