Anti-PGM1 antibody [EPR15241]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-PGM1 antibody [EPR15241] (AB192876)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% tritonX-100 permeabilized HeLa cells labeling PGM1 with ab192876 at 1/500 dilution followed by Goat anti rabbit IgG (AlexaFluor® 488) (ab150077) secondary antibody at 1/400 dilution. Nuclear counter stain is DAPI (blue).

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-PGM1 antibody [EPR15241] (AB192876)

ab192876 staining PGM1 in A549 (human lung carcinoma) by intracellular flow cytometry. Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. The sample was incubated with the primary antibody at a dilution of 1/90. A goat anti rabbit IgG (Alexa Fluor^® 488) at a dilution of 1/2000 was used as the secondary antibody.

Isoytype control : Rabbit monoclonal IgG (Black)

Unlabelled control : Cell without incubation with primary antibody and secondary antibody (Blue)

• IP

Supplier Data

Immunoprecipitation - Anti-PGM1 antibody [EPR15241] (AB192876)

Western blot analysis of PGM1 immunoprecipitated from Human fetal liver lysate with ab192876 at 1/60 dilution. Lane 1 : Human fetal liver lysate. Lane 2 : PBS instead of Human fetal liver lysate.
Secondary : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1500 dilution.

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-PGM1 antibody [EPR15241] (ab192876)

Predicted band size: 61 kDa

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• WB

Supplier Data

Western blot - Anti-PGM1 antibody [EPR15241] (AB192876)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-PGM1 antibody [EPR15241] (ab192876) at 1/10000 dilution

Lane 1:

HepG2 cell lysate at 20 µg

Lane 2:

293T cell lysate at 20 µg

Lane 3:

Human fetal liver lysate at 20 µg

Lane 4:

Human fetal heart lysate at 20 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

Predicted band size: 61 kDa

Observed band size: 61 kDa

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• WB

Supplier Data

Western blot - Anti-PGM1 antibody [EPR15241] (AB192876)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-PGM1 antibody [EPR15241] (ab192876) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 10 µg

Lane 2:

Mouse heart tissue lysate at 10 µg

Lane 3:

Mouse kidney tissue lysate at 10 µg

Lane 4:

Mouse spleen tissue lysate at 10 µg

Lane 5:

Rat kidney tissue lysate at 10 µg

Lane 6:

Rat spleen tissue lysate at 10 µg

Lane 7:

C6 cell lysate at 10 µg

Lane 8:

RAW 264.7 cell lysate at 10 µg

Lane 9:

PC12 cell lysate at 10 µg

Lane 10:

NIH/3T3 cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugate at 1/1000 dilution

Predicted band size: 61 kDa

Observed band size: 61 kDa

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• WB

CiteAb

Western blot - Anti-PGM1 antibody [EPR15241] (AB192876)

PGM1 western blot using anti-PGM1 antibody [EPR15241] ab192876. Publication image and figure legend from Ma, J., Wei, K., et al., 2020, Nat Commun, PubMed 32286295.

ab192876 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 ab192876 please see the product overview.

Glycogen is synthesized in inflammatory macrophages.a–c Intracellular glycogen levels in untreated, IFN-γ/LPS (20 ng mL-1 IFN-γ plus 100 ng mL-1 LPS) or IL-4 (10 ng mL-1) treated BMDMs were detected and observed by PAS staining (scale bar, 20 μm) (a), colorimetric assay (b) and TEM (scale bar, 0.5 μm) (c). The arrows point to intracellular glycogen deposits. d, e Pgm1, Ugp2, and Gys1 expression in untreated, IFN-γ/LPS or IL-4 treated BMDMs were determined by real-time PCR (d) and western blot (e). f Overview of three glucose metabolic pathways : glycogen metabolism (left), glycolysis (middle) and PPP (right) and the inhibitors (GPI/6AN) are shown. g–i 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 + 6-labeled G6P/G1P (g), m + 6-labeled UDPG (h) and m + 3-labeled G6P/G1P (i). j, k Pck1, Fbp1, and G6pase expression in untreated, IFN-γ/LPS or IL-4 treated BMDMs were determined by real-time PCR (j) and western blot (k). l Consumption of glucose in untreated, IFN-γ/LPS or IL-4 treated BMDMs were measured by enzymatic methods. m Relative mRNA expression of Slc2a1/2 and Hk1/2/3 in untreated, IFN-γ/LPS or IL-4 treated BMDMs were determined by real-time PCR. n, oHk1/2/3, Pgm1 or Gys1 siRNA transfected BMDMs were stimulated with IFN-γ/LPS for 36 h. Intracellular glycogen levels were detected by colorimetric assay. Unless otherwise specified, n = 3 biologically independent experiments were performed. Data are presented as mean ± SEM. P values were calculated using one-way ANOVA, ****p < 0.0001.

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