The Flag-CTL and Flag-GDPD5 plasmids were transfected into SH-SY5Y cells with Neofect™ DNA transfection reagent. After cells were transfected for 48 h, the cells were lysed by 1% SDS lysing buffer containing Protease Inhibitor Cocktail and Phosphatase Inhibitor Cocktail (Apexbio, Houston, TX, USA) for Western blot analysis. The protein concentration was determined by a BCA protein assay reagent kit (Thermo Scientific, Waltham, MA, USA). All blots were, respectively, incubated with primary anti-bodies anti-flag (1:1000, ABclonal, Wuhan, China), anti-p-ACC, anti-ACC, Anti-p-ACLY, anti-ACLY, anti-PFKFB3, anti-HK2 and anti-LDHA (1:1000, Cell Signaling Technology, MA, USA), anti-ALDOA, anti-ENO2, anti-HADH and anti-PPARA (1:1000, proteintech, Wuhan, China), as well as anti-β-Tubulin (1:5000, TRANSGEN, Beijing, China). Bands were visualized with ECL Reagents (Smart-Lifesciences, Changzhou, China).
Anti pfkfb3
Manufactured by Cell Signaling Technology
Sourced in China
Anti-PFKFB3 is a primary antibody that recognizes the PFKFB3 protein. PFKFB3 is an enzyme that plays a key role in glycolysis, the metabolic pathway responsible for converting glucose into energy. This antibody can be used for the detection and analysis of PFKFB3 in various experimental applications.
Automatically generated - may contain errors
Lab products found in correlation
Bca protein assay reagent kit, by Thermo Fisher Scientific (2 mentions)
Anti p acc, by Cell Signaling Technology (2 mentions)
Anti acc, by Cell Signaling Technology (2 mentions)
Anti hk2, by Cell Signaling Technology (2 mentions)
Protease inhibitor cocktail and phosphatase inhibitor cocktail, by Apexbio (2 mentions)
Anti aldoa, by Proteintech (2 mentions)
Anti β tubulin, by Transgene (2 mentions)
Anti ldha, by Cell Signaling Technology (2 mentions)
Anti acly, by Cell Signaling Technology (2 mentions)
Anti p acly, by Cell Signaling Technology (2 mentions)
Anti eno2, by Proteintech (2 mentions)
Anti hadh, by Proteintech (2 mentions)
Anti ppara, by Proteintech (2 mentions)
Ecl reagents, by Smart-Lifesciences (2 mentions)
Anti flag, by ABclonal (2 mentions)
Protease and phosphatase inhibitor, by Roche (1 mentions)
Enhanced chemi luminescence (ecl), by Thermo Fisher Scientific (1 mentions)
Cell lysis buffer, by Cell Signaling Technology (1 mentions)
Anti histone h3, by Cell Signaling Technology (1 mentions)
Anti α tubulin, by Merck Group (1 mentions)
3 protocols using anti pfkfb3
1
Flag-Tagged Protein Expression Analysis
2
Investigating Metabolic Regulators in OSRC2 Cells
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The GFP-CTL and GFP-HMGCS2 plasmids were transfected into OSRC2 cells with Neofect™ DNA transfection reagent. After cells were transfected for 48 h, the cells were lysed by 1% SDS lysing buffer containing Protease Inhibitor Cocktail and Phosphatase Inhibitor Cocktail (Apexbio, Houston, TX, USA) for Western blot analysis. The protein concentration was determined by a BCA protein assay reagent kit (Thermo Scientific, Waltham, MA, USA). All blots were, respectively, incubated with primary anti-bodies anti-flag (1:1000, ABclonal, Wuhan, China), anti-p-ACC, anti-ACC, Anti-p-ACLY, anti-ACLY, anti-PFKFB3, anti-HK2 and anti-LDHA (1:1000, Cell Signaling Technology, MA, USA), anti-ALDOA, anti-ENO2, anti-HADH and anti-PPARA (1:1000, proteintech, Wuhan, China), and anti-β-Tubulin (1:5000, TRANSGEN, Beijing, China). Bands were visualized with ECL Reagents (Smart-Lifesciences, Changzhou, China).
3
Immunoblotting Protein Analysis Protocol
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Protein extraction and immunoblot analysis were performed using a modified Laemmli sample buffer (125 mM Tris-HCl, pH 6.8 buffer containing 2% SDS and 10% glycerol) or cell lysis buffer (Cell Signaling Technology) in the presence of protease and phosphatase inhibitors (Roche). Lysates were separated by SDS-PAGE under reducing conditions, transferred to a nitrocellulose or PVDF membrane, and analyzed by immunoblotting. Primary antibodies used were rabbit anti-CPT1A (Cell Signaling Technology), anti-total AMPKa (Cell Signaling Technology), rabbit anti-phospho-AMPKa Thr172 (Cell Signaling Technology), anti-PFKFB3 (Cell Signaling Technology), anti-NICD (Cell Signaling Technology), anti-Histone H2A (Cell Signaling Technology), anti-Acetyl-Histone H2A (Lys5) (Cell Signaling Technology), anti-Histone H3 (Cell Signaling Technology), anti-Acetyl-Histone H3 (Lys9) (Cell Signaling Technology), anti-Histone H4 (Cell Signaling Technology), anti-Acetyl-Histone H4 (Lys8) (Cell Signaling Technology), anti-a-tubulin (Sigma) and anti-acetyllysine (PTM-Biolab). Appropriate secondary antibodies were from Cell Signaling Technology. Signal was detected using the ECL or Femto system (Thermo Fisher Scientific) according to the manufacturer's instructions. Densitometric quantifications of bands were done with Fiji software (https://fiji.sc).
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