Characterization
Description(s):The peroxisome-proliferator activated receptor gamma (PPARgamma) is a member of the nuclear receptor superfamily of transcription factors. PPARgamma is important to a variety of biological processes, such as adipocyte differentiation, glucose homeostasis, lipid trafficking as well as vascular function and hypertension. PPARgamma exerts its effects by regulating target gene transcription in a ligand-dependent manner. Upon activation, the PPARgamma-RXR heterodimer binds to specific DNA sequences on the target gene promoter and leads to transcriptional regulation.
Signosis has developed a plate-based array for profiling 20+ PPARgamma-regulated genes. By using this assay, researchers are able to compare gene expression in up to three samples on a single microtiter plate.
Applicable Grid:
List of Applicable Genes
| 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | |
| A | PKA (Prkaca) | NfkB (NfkB1) | CD36 | PKA (Prkaca) | NfkB (NfkB1) | CD36 | PKA (Prkaca) | NfkB (NfkB1) | CD36 | PKA (Prkaca) | NfkB (NfkB1) | CD36 |
| B | PKC (Prkcg) | NOS1 | CD40 | PKC (Prkcg) | NOS1 | CD40 | PKC (Prkcg) | NOS1 | CD40 | PKC (Prkcg) | NOS1 | CD40 |
| C | cAMP | NOS2 | B-actin | cAMP | NOS2 | B-actin | cAMP | NOS2 | B-actin | cAMP | NOS2 | B-actin |
| D | ERK (EphB1) | NOS3 | Klotho | ERK (EphB1) | NOS3 | Klotho | ERK (EphB1) | NOS3 | Klotho | ERK (EphB1) | NOS3 | Klotho |
| E | JNK (Mapk8) | PPARa | Nephrin (Nphs1) | JNK (Mapk8) | PPARa | Nephrin (Nphs1) | JNK (Mapk8) | PPARa | Nephrin (Nphs1) | JNK (Mapk8) | PPARa | Nephrin (Nphs1) |
| F | Raf-1 | PPARß/d | Rho | Raf-1 | PPARß/d | Rho | Raf-1 | PPARß/d | Rho | Raf-1 | PPARß/d | Rho |
| G | GRK-2 (Adrbk1) | PPARgamma | GAPDH | GRK-2 (Adrbk1) | PPARgamma | GAPDH | GRK-2 (Adrbk1) | PPARgamma | GAPDH | GRK-2 (Adrbk1) | PPARr | GAPDH |
| H | GRK-4 | NFAT (Nfat5) | Blank | GRK-4 | NFAT (Nfat5) | Blank | GRK-4 | NFAT (Nfat5) | Blank | GRK-4 | NFAT (Nfat5) | Blank |
Principle
Signosis’ proprietary cDNA plate array is a plate-based hybridization profiling analysis for monitoring the expression of dozens of genes through reverse transcription of mRNA into cDNA. Like array analyses, total RNA is first reverse transcribed into cDNA in the presence of biotin-dUTP in the assay. Targeted genes are then specifically captured onto individual wells on a plate, instead of membranes, through a pre-coated gene-specific oligonucleotide. The captured cDNAs are further detected with streptavidin-HRP. Luminescence is reported as relative light units (RLUs) on a microplate luminometer. The expression level of genes is directly proportional to the luminescent intensity.
