Rat phosphoenolpyruvate carboxykinase,PCK ELISA Kit

Instructions
Code CSB-E13612r
Size 96T,5×96T,10×96T
See More Details 24T ELISA kits trial application
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Product Details

Target Name phosphoenolpyruvate carboxykinase 1 (soluble)
Alternative Names Pck1 ELISA Kit; Phosphoenolpyruvate carboxykinase ELISA Kit; cytosolic [GTP] ELISA Kit; PEPCK-C ELISA Kit; EC 4.1.1.32 ELISA Kit
Abbreviation PCK1
Uniprot No. P07379
Species Rattus norvegicus (Rat)
Sample Types serum, plasma, tissue homogenates, cell lysates
Detection Range 25 pg/mL-1600 pg/mL
Sensitivity 6.25 pg/mL
Assay Time 1-5h
Sample Volume 50-100ul
Detection Wavelength 450 nm
Research Area Metabolism
Assay Principle quantitative
Measurement Sandwich
Precision
Intra-assay Precision (Precision within an assay): CV%<8%        
Three samples of known concentration were tested twenty times on one plate to assess.    
Inter-assay Precision (Precision between assays): CV%<10%        
Three samples of known concentration were tested in twenty assays to assess.      
               
Linearity
To assess the linearity of the assay, samples were spiked with high concentrations of rat PCK in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay.  
  Sample Serum(n=4)    
1:1 Average % 94    
Range % 87-98    
1:2 Average % 87    
Range % 83-92    
1:4 Average % 91    
Range % 85-97    
1:8 Average % 107    
Range % 102-111    
Recovery
The recovery of rat PCK spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section.  
 
Sample Type Average % Recovery Range    
Serum (n=5) 95 90-99    
EDTA plasma (n=4) 104 97-108    
               
               
Typical Data
These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.  
 
pg/ml OD1 OD2 Average Corrected    
1600 2.543 2.468 2.506 2.367    
800 1.742 1.656 1.699 1.560    
400 1.076 1.033 1.055 0.916    
200 0.612 0.591 0.602 0.463    
100 0.376 0.380 0.378 0.239    
50 0.283 0.278 0.281 0.142    
25 0.209 0.205 0.207 0.068    
0 0.140 0.138 0.139      
Troubleshooting
and FAQs
ELISA kit FAQs
Storage Store at 2-8°C. Please refer to protocol.
Lead Time 3-5 working days

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Target Data

Function Catalyzes the conversion of oxaloacetate (OAA) to phosphoenolpyruvate (PEP), the rate-limiting step in the metabolic pathway that produces glucose from lactate and other precursors derived from the citric acid cycle.
Gene References into Functions
  1. Peroxynitrite-induced coronary vascular dysfunction may be mediated, at least in part, through increased expressions and activities of RhoA, ROCK1, and ROCK2. PMID: 27724862
  2. GBS can reduce blood glucose in T2DM rats while improving glucose tolerance and hyperglycemia, and the mechanism appears to be associated with a decrease of hepatic PEPCK mRNA and protein expression. PMID: 26681041
  3. PEPCK is downregulated and has a role in reduction of blood glucose and gluconeogenesis after phenobarbital administration in rats PMID: 26348778
  4. It observed lower expression of hepatic PEPCK mRNA, in the magnesium-deficient rats, thus suggesting a possible compensatory mechanism to diminish glycogenesis. A low-magnesium diet alters glucocorticoid metabolism, which leads to endothelial damage. PMID: 23015202
  5. Leptin activates NOS III and induces nitric oxide that nitrates PEPCK-C to reduce its level and glyceroneogenesis. PMID: 22808220
  6. The increase of protein intake during pregnancy could decrease the expression of y acid synthetase and PEPCK mRNA, and prevent offspring from high-fat diet-induced obesity in adult. PMID: 19548567
  7. retinoic acid response element 2 in rat Pck1 promoter is responsible for transcriptional activation by retinoic acid PMID: 21519922
  8. In addition to increased transcription rate, a gestational high fat diet resulted in modifications of the Pck1 histone code in livers of offspring. PMID: 21486814
  9. Pck1 plays an important role in both the mammary gland adipocytes and epithelial cells during lactation PMID: 21504969
  10. During acute hepatic failure severe endotoxemia damages the function of gluconeogenesis in livers and kidneys by inhibiting transcription of PEPCK. PMID: 17903369
  11. Taken together, our results suggest that, during very demanding exercise, muscle-derived interleukin-6 could help increase hepatic glucose production by directly upregulating PEPCK mRNA abundance. PMID: 19850730
  12. Expression of the PEPCK gene in H4IIEC3 rat hepatoma cells is closely coordinated with cell proliferation. PMID: 11820793
  13. increased PEPCK gene transcription at acid pH required a cis-acting element (enhancer) in the more distal 5' flanking region of the promoter PMID: 11939717
  14. polyunsaturated and nonmetabolized FAs stimulate adipose PEPCK, therefore potentially enhancing glyceroneogenesis and reducing FA output. This mechanism could participate in the hypolipidemic action of PUFAs. PMID: 11968005
  15. promoter context may influence which domains within a transcription factor are employed to mediate transactivation of PEPCK PMID: 12237288
  16. transcription of the gene for phosphoenolpyruvate carboxykinase (GTP) in the liver is regulated by SREBP-1c and Sp1 PMID: 14744869
  17. PEPCK transcription induced by protein kinase A is regulated by CREB and C/EBP. PMID: 15604115
  18. SHARP-2 is a potential repressor of PEPCK gene expression PMID: 15733865
  19. Hepatic PEPCK gene expression is modulated by chronic exercise training. PMID: 16458327
  20. Pck1 is up-regulated during liver gluconeogenesis. PMID: 17158265
  21. Endogenous lipid molecules induce PEPCK-C gene transcription and attenuate insulin action in liver. PMID: 17678617
  22. Comparison of the Mn2+-OAA-Mn2+GDP and Mn2+-Mn2+GTP structures illustrates a marked difference in the bound conformations of the nucleotide substrates in which the GTP nucleotide is bound in a high-energy state. PMID: 17685635
  23. Suppression of phosphoenolpyruvate carboxykinase gene expression by reduced endogenous glutathione level PMID: 17964299
  24. The effects of several variants of retinoic acid on the expression and activity of PEPCK in human, mouse, and rat adipose tissue are reported. PMID: 18492826
  25. structural data are consistent with a model in that the energy input from substrate association results in changes in the free energy landscape for the protein, allowing for structural transitions along an induced fit pathway PMID: 18772387
  26. skeletal muscle PEPCK has a role in determining physical activity levels PMID: 19521512
  27. Expression of the rat PEPCK gene in adipose tissue requires a PPAR-gamma/RXR binding site located 1 kbp upstream of the promoter. Similar elements are in the human and mouse gene. PMID: 10224131
  28. In white adipose tissue, overexpression causes obesity without diabetes, while underexpression causes lipodystrophy and perhaps insulin resistance. PMID: 12409311
  29. PEPCK catalyzes a key step of gluconeogenesis and glyceroneogenesis. It is expressed at significant levels in liver, kidney, intestine, white and brown adipose tissues, and lactating mammary gland. Control is mainly at transcription. PMID: 9242918
  30. PEPCK is a key enzyme of glyceroneogenesis in adipocytes and hepatocytes. Glyceroneogenesis produces glycerol-3-P to control the triglyceride/fatty acid cycle. PMID: 12788931
  31. The first cAMP response element (CRE) was discovered upstream of the rat PEPCK gene. PMID: 3015903
  32. Expression of the rat PEPCK gene in adipose tissue requires a PPAR-gamma/RXR binding site located 1 kbp upstream of the promoter. Similar elements are in the human and mouse gene. PMID: 10224131

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Subcellular Location Cytoplasm
Protein Families Phosphoenolpyruvate carboxykinase [GTP] family
Database Links

KEGG: rno:362282

STRING: 10116.ENSRNOP00000030913

UniGene: Rn.104376

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