Tumors were fixed in formalin for 24 h, paraffin embedded, sectioned, and stained according to standard procedures. Briefly, endogenous peroxidases were inactivated by 3% hydrogen 673 peroxide. Non-specific signals were blocked using 3% BSA, 10% goat serum in 0.1% Triton X-100. After antigen retrieval in citrate buffer, slides were stained using respective antibodies overnight at 4°C, [ ENO1 (Proteintech, #11204–1-AP), PGK1 (Proteintech #17811–1-AP), PGAM1 (Proteintech, #16126–1-AP), Tyrosinase (Abcam, ab738), pAKT (Cell Signaling, #9271), pIGF1R (Abcam, ab39398)]. After overnight incubation, the slides were washed and incubated with secondary antibody (HRP-polymers, Biocare Medical) for 30 min at room temperature. The slides were washed three times and stained with DAB substrate (ThermoFisher Scientific). The slides were then counterstained with hematoxylin and mounted with mounting medium.
Pgam1
Manufactured by Proteintech
Sourced in China
PGAM1 is a glycolytic enzyme that catalyzes the conversion of 3-phosphoglycerate to 2-phosphoglycerate in the glycolytic pathway. It plays a key role in energy production through the breakdown of glucose.
Automatically generated - may contain errors
Lab products found in correlation
Hrp polymers, by Biocare Medical (2 mentions)
Pigf1r, by Abcam (2 mentions)
P akt, by Cell Signaling Technology (2 mentions)
Dab substrate, by Thermo Fisher Scientific (2 mentions)
Tyrosinase, by Abcam (2 mentions)
Imagequant las 4000 mini, by GE Healthcare (1 mentions)
Cell lysis buffer, by Beyotime (1 mentions)
Enhanced bca protein assay kit, by Beyotime (1 mentions)
Ecl reagent, by GE Healthcare (1 mentions)
Gapdh, by Beyotime (1 mentions)
3 protocols using pgam1
1
Immunohistochemical Analysis of Tumor Markers
2
Immunohistochemical Analysis of Tumor Markers
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Tumors were fixed in formalin for 24 h, paraffin embedded, sectioned, and stained according to standard procedures. Briefly, endogenous peroxidases were inactivated by 3% hydrogen 673 peroxide. Non-specific signals were blocked using 3% BSA, 10% goat serum in 0.1% Triton X-100. After antigen retrieval in citrate buffer, slides were stained using respective antibodies overnight at 4°C, [ ENO1 (Proteintech, #11204–1-AP), PGK1 (Proteintech #17811–1-AP), PGAM1 (Proteintech, #16126–1-AP), Tyrosinase (Abcam, ab738), pAKT (Cell Signaling, #9271), pIGF1R (Abcam, ab39398)]. After overnight incubation, the slides were washed and incubated with secondary antibody (HRP-polymers, Biocare Medical) for 30 min at room temperature. The slides were washed three times and stained with DAB substrate (ThermoFisher Scientific). The slides were then counterstained with hematoxylin and mounted with mounting medium.
3
Western Blot Protein Analysis Protocol
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The Western blot analysis was performed as described previously (15 (link)). Briefly, the cell and tissue sample lysates, extracted by cell lysis buffer (Beyotime, China) and the concentration was measured by Enhanced BCA Protein Assay Kit (Beyotime, China) according to the manufacturer’s instructions. About 20 μg of protein was separated by 12% SDS-PAGE and transferred to a nitrocellulose membrane, respectively. Then the membrane was blocked with 5% bovine serum albumin (BSA) in Tris-buffered saline with Tween (TBST) for 2 h at room temperature, followed by incubated overnight with primary antibodies against PGAM1 (1:500 dilution, Proteintech, China), GAPDH (1:1000 dilution, Beyotime, China). After incubating with the first antibody, the membrane was washed three times with TBST, and then incubated with the relevant secondary antibodies (1:6000 dilution, Promega, USA) for 1 h at room temperature and followed by three times washes, then visualized by using the ECL reagent (GE Healthcare, UK), the protein bands were subsequently measured using the ImageQuant LAS 4000mini (GE Healthcare, USA) and grayscale analysis using the ImageJ 1.42q (ImageJ software, Way Rasband, National Institutes of Health, USA).
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