Triglyceride concentrations were measured using GPO-PAP micro-test (purchased from Roche, Mannheim, Germany). Liver cholesterol was determined by an assay from Diaglobal (Berlin, Germany). Serum lipids, hepatic sphingomyelin and ceramide were quantified by direct flow injection electrospray ionization tandem mass spectrometry (ESI-MS/MS) in positive ion mode using the analytical setup and strategy as described previously [13 (link), 14 (link)].
Gpo pap micro test
Manufactured by Roche
The GPO-PAP micro-test is a laboratory equipment product designed for the quantitative determination of total glycerol in various biological samples. It utilizes an enzymatic method based on the glycerol-3-phosphate oxidase and peroxidase (GPO-PAP) principle to measure the glycerol concentration.
Automatically generated - may contain errors
Lab products found in correlation
Quantichrom glucose assay kit, by Biotrend (1 mentions)
Camp elisa, by Enzo Life Sciences (1 mentions)
X tremegene transfection reagent, by Roche (1 mentions)
Endo porter, by Gene Tools (1 mentions)
Insulin elisa, by Mercodia (1 mentions)
3t3 l1 preadipocyte, by American Type Culture Collection (1 mentions)
3 protocols using gpo pap micro test
1
Lipid Quantification Protocol
2
Metabolic Biomarker Quantification
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Triglyceride concentrations were measured using GPO-PAP microtest (purchased from Roche, Mannheim, Germany) and total cholesterol in serum was determined by using an assay from Diaglobal (Berlin, Germany). Proinsulin and insulin were determined by the appropriate ELISAs from Mercodia (Uppsala, Sweden). Glucose was measured by QuantiChrom Glucose Assay Kit from Biotrend (Köln, Germany). The Homeostasis model assessment (HOMA) index was calculated using the formula: [fasting glucose (mmol/L) × fasting insulin (mU/L)]/22.5.
3
Glucose, Lipid, and Gene Expression Analysis in 3T3-L1 Adipocytes
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QuantiChrom TM Glucose Assay Kit was from Biotrend (Köln, Germany). IP/Co-IP Kit Pierce Protein A/G Magnetic was from Thermo Scientific (Schwerte, Germany). Triglyceride concentrations were measured using GPO-PAP micro-test (Roche). Free fatty acids, glycerol and cholesterol were determined by assays from BioCat. The cAMP ELISA was from EnzoLife Science (Lörrach, Germany). Insulin ELISA was from Mercodia (Uppsala, Sweden).
Cell culture and siRNA transfection 3T3-L1 preadipocytes were from the American Type Culture Collection (ATCC, Manassas, VA, USA) and differentiated as described [33] . Transfection of preadipocytes with siRNAs was performed with XtremeGene transfection reagent (Roche, Mannheim, Germany) or Endoporter (Gene Tools LLC, Philomath, Oregon, USA) [12] (link). Differentiated adipocytes were transfected by electroporation as described [12] (link). Sequences of the siRNAs used have been published earlier [12] (link).
Immunoblot and real-time RT-PCR Immunoblot and real-time RT-PCR analysis were performed as described [34, 35] . 18S rRNA was used for normalization of gene expression with the exception of data related to patients were GAPDH was used [31] . Primers and antibodies used are listed in table 1 and2
Cell culture and siRNA transfection 3T3-L1 preadipocytes were from the American Type Culture Collection (ATCC, Manassas, VA, USA) and differentiated as described [33] . Transfection of preadipocytes with siRNAs was performed with XtremeGene transfection reagent (Roche, Mannheim, Germany) or Endoporter (Gene Tools LLC, Philomath, Oregon, USA) [12] (link). Differentiated adipocytes were transfected by electroporation as described [12] (link). Sequences of the siRNAs used have been published earlier [12] (link).
Immunoblot and real-time RT-PCR Immunoblot and real-time RT-PCR analysis were performed as described [34, 35] . 18S rRNA was used for normalization of gene expression with the exception of data related to patients were GAPDH was used [31] . Primers and antibodies used are listed in table 1 and2
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