Tissue samples were embedded in paraffin and stained using hematoxylin and eosin (HE) used for immunohistochemistry (performed by Morphotechnology Co. Ltd., Sapporo, Japan) using primary antibodies against Pygl (diluted 1:75, Cat# Ab198268, Abcam, Cambridge, UK) or osteocalcin (diluted 1:1,000, Cat# Ab198268, Abcam), for 1 h at room temperature, according to the polymer method. Periodic acid-Schiff (PAS) staining, along with amylase digestion, was performed as previously described [19 (link)].
Ab198268
Manufactured by Abcam
Sourced in United Kingdom
Ab198268 is a laboratory product manufactured by Abcam. It is a reagent used for research purposes.
Automatically generated - may contain errors
Lab products found in correlation
Ripa lysis buffer, by Beyotime (1 mentions)
Ecl kit, by Abcam (1 mentions)
Ab92494, by Abcam (1 mentions)
Ab9483, by Abcam (1 mentions)
Ab52492, by Abcam (1 mentions)
Ab109250, by Abcam (1 mentions)
Ab222782, by Abcam (1 mentions)
Ab200834, by Abcam (1 mentions)
Ab154969, by Abcam (1 mentions)
Ab227043, by Abcam (1 mentions)
Ab128870, by Abcam (1 mentions)
Ab174830, by Abcam (1 mentions)
Ab234111, by Abcam (1 mentions)
Ab137043, by Abcam (1 mentions)
4 protocols using ab198268
1
Histological Examination of Tissue Samples
2
Western Blot Analysis of Stem Cell Markers
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Total proteins of ESCA cells was extracted by radio-immunoprecipitation assay (RIPA) lysis buffer (Beyotime) [32 (link)]. Subsequently, equal amounts of protein were transferred into polyvinylidene difluoride (PVDF) membranes after separation by 10% sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS-PAGE). The membranes were blocked with 5% nonfat milk and then incubated with primary antibodies against CD133 (ab222782, 1:2000, Abcam), Nanog (ab109250, 1:1000, Abcam), Oct4 (ab200834, 1:10,000, Abcam), Sox2 (ab92494, 1:1000, Abcam), ALDH1 (ab52492, 1:2000, Abcam), PYGL (ab198268, 1:1000, Abcam) and GAPDH (ab9483, 1:1000, Abcam) overnight at 4°C. GAPDH was regarded as the internal reference. Subsequently, the membranes were washed with Tris-buffered saline Tween-20 (TBST), followed by incubation with horseradish peroxidase (HRP)‐conjugated secondary antibodies for 1 h at room temperature. Finally, the enhanced chemiluminescence (ECL) kits (Abcam) were used to visualize the protein bands.
3
Glycogen Metabolism Profiling in Echinococcus granulosus
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Echinococcus granulosus PSCs collected from liver hydatid cysts of newly slaughtered sheep were homogenized with lysis buffer at 4°C and centrifuged at 8000 × g for 15 min. Then, 20 μg of the protein-containing supernatant was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and the Polyvinylidene fluoride (PVDF) membrane was blocked for 1 h and incubated with anti-PYGB (Abcam, United Kingdom, ab154969), anti-PYGL (Abcam, ab198268) and anti-PYGM (Abcam, ab231963) primary antibodies (1/1,000) at 4°C overnight. Then, the membrane was incubated with a 1/5,000 dilution of a goat anti-rabbit IgG secondary antibody conjugated with HRP (Abcam, United Kingdom, ab6721) at room temperature for 1 h before washed again. An ECL Western Blotting Substrate Kit (Tanon, China) was used to detect the proteins on the PVDF membrane.
4
Protein Extraction and Western Blot Analysis
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The frozen tissues were homogenized in a lysis buffer (20 mM Tris-HCl, 150 mM NaCl, 1 mM EDTA, 1 mM EGTA, 1% Triton-X100, and protease inhibitor, pH 7.4) and then centrifuged for 20 min at 10,000 g to discard cell debris. The total protein concentrations were determined using a Bio-Rad kit. The proteins were subjected to Western blot analysis with the desired antibodies. Antibodies against carnitine palmitoyltransferase 1A (CPT1A) (ab234111), HMGCS2 (ab137043), FAS (ab128870), HMGCR (ab174830), liver glycogen phosphorylase (PYGL) (ab198268) and PYGL (phospho S15, ab227043) were obtained from Abcam. Antibodies against COX4 (A21348) was obtained from Invitrogen. Antibody against phosphoenolpyruvate carboxykinase 1 (PCK1) (A2036) and GCK (A6293) was purchased from ABclonal. Antibodies against AMPKα (2532), phospho-AMPKα (2535), CREB (9197), Phospho-CREB (9198), and SCD1 (2438) were purchased from Cell Signaling Technology. Antibody against G6PC (22169-1-AP) was purchased from Proteintech. Antibody against TUBULIN (T0198) was obtained from Sigma-Aldrich.
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