PPAR alpha/NR1C1 Overexpression Lysate Summary
| Description |
PPAR alpha/NR1C1 Transient Overexpression Lysate
Expression Host: HEK293T
Plasmid: RC216237
Accession#: NM_001001928
Protein Tag: C-MYC/DDK
You will receive 1 vial of lysate (100ug), 1 vial of empty vector negative control (100ug), and 1 vial of 2xSDS sample buffer (250ul). Each vial of cell lysate contains 100ug of total protein (at 1 mg/ml). The 2xSDS Sample Buffer consists of 4% SDS, 125mM Tris-HCl pH6.8, 10% Glycerol, 0.002% Bromophenol blue, 100mM DTT. |
| Gene |
PPARA |
Applications/Dilutions
| Dilutions |
|
| Application Notes |
This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag ( NBP1-71705) present on the protein construct. Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading. |
| Theoretical MW |
52 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors. |
Packaging, Storage & Formulations
| Storage |
Store at -80C. Avoid freeze-thaw cycles. |
| Buffer |
RIPA buffer |
Lysate Details for Array
Notes
HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.
Alternate Names for PPAR alpha/NR1C1 Overexpression Lysate
Background
Peroxisome proliferators are non-genotoxic carcinogens which are purported to exert their effect on cells through their interaction with members of the nuclear hormone receptor family, termed peroxisome proliferator activated receptors (PPARs). Nuclear hormone receptors are ligand-dependent intracellular proteins that stimulate transcription of specific genes by binding to specific DNA sequences following activation by the appropriate ligand. Studies indicate that PPARs are activated by peroxisome proliferators such as clofibric acid, nafenopin, and WY-14,643, as well as by some fatty acids. It has also been shown that PPARs can induce transcription of acyl coenzyme A oxidase & cytochrome P450 A6 (CYP450 A6) through interaction with specific response elements. PPAR alpha is activated by free fatty acids including linoleic, arachidonic, and oleic acids. Induction of peroxisomes by this mechanism leads to a reduction in blood triglyceride levels. PPAR alpha is expressed mainly in skeletal muscle, heart, liver, and kidney and is thought to regulate many genes involved in the beta-oxidation of fatty acids. Activation of rat liver PPAR alpha has been shown to suppress hepatocyte apoptosis. PPAR alpha, like several other nuclear hormone receptors, heterodimerizes with retinoic X receptor (RXR) alpha to form a transcriptionally competent complex. The corresponding gene for the PPAR alpha is NR1C1.
Limitations
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are
guaranteed for 6 months from date of receipt.
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Publications for PPAR alpha/NR1C1 Lysate (NBP2-10726) (0)
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Product General Protocols
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Video Protocols
FAQs for PPAR alpha/NR1C1 Lysate (NBP2-10726). (Showing 1 - 1 of 1 FAQ).
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Please differentiate to me between PPAR and PGC clearly. I am confused with the difference between these two
- Thank you very much for contacting Novus Biologicals technical support team and sharing your query on the differences between PGC-1 alpha and PPAR. These are two different proteins encoded by their respective genes and serves different functions. PGC-1 alpha (PGC1A or PPAR gamma coactivator 1-alpha) is a transcriptional co-activator for steroid receptors and nuclear receptors, and it regulates diverse aspects of cellular physiology. It up-regulates the transcriptional activity of PPAR-gamma /thyroid hormone receptor on the uncoupling protein promoter; regulates the key mitochondrial genes involved in adaptive thermogenesis; implicates in the metabolic reprogramming in response to nutrients availability through the coordination of the expression of a wide array of genes involved in the regulation of glucose and fatty acid metabolism. Among our PGC-1 alpha antibodies, NBP1-04676 is our best selling product with nice customer feedback and citations in at least 13 research publications. PPAR (PPAR alpha) on the other hand is a ligand-activated transcription factor which gets activated by the endogenous ligand 1-palmitoyl-2-oleoyl-sn-glycerol-3-phosphocholine, and oleylethanolamide (a naturally occurring lipid that regulates satiety), and acts as a key regulator of lipid metabolism. It also acts as a receptor for peroxisome proliferators such as hypolipidemic drugs and fatty acids. It regulates the peroxisomal beta-oxidation pathway of fatty acids, and also functions as transcription activator for the ACOX1 and P450 genes. We have a variety of PPAR alpha antibodies. I hope you will find this information helpful but please let me know if I can support you with anything else from my end. Thank you very much for choosing Novus Biologicals as your quality reagent supplier and we wish you the best with your research projects.
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