PLA2G4C Overexpression Lysate Summary
| Description |
PLA2G4C Transient Overexpression Lysate Expression Host: HEK293T
Plasmid: RC209567
Accession#: NM_003706
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 |
PLA2G4C |
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 |
60.8 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 PLA2G4C Overexpression Lysate
Background
PLA2G4C, also known as Cytosolic phospholipase A2 gamma, has 3 isoforms, a 541 amino acid isoform that is 61 kDa, a 527 amino acid isoform that is 59 kDa and a 551 amino acid isoform that is 62 kDa; highly expressed in heart and skeletal muscle; hydrolyzes glycerophospholipids to produce free fatty acids and lysophospholipids, both of which serve as precursors in the production of signaling molecules has been shown to be a calcium-independent and membrane bound enzyme, and has a preference for arachidonic acid at the sn-2 position of phosphatidylcholine as compared with palmitic acid. This protein has being studied for its involvement in oligodendroglioma, ovarian cancer, breast cancer, schizophrenia, and neuronitis. PLA2G4C protein has been observed with relation to S100A10, MBOAT2, GPCPD1, LPCAT1, and LPCAT2 in immune response CD16 signaling in NK cells, development Leptin signaling via JAK/STAT and MAPK cascades, development Alpha-2 adrenergic receptor activation of ERK, immune response CCR3 signaling in eosinophils, neurophysiological process PGE2-induced pain processing, chemokine signaling, DHA signaling, VEGF family ligands and receptor interactions, antioxidant action of vitamin-C, LDL oxidation in atherogenesis, phospholipases, acyl chain remodelling of PE, metabolism, acyl chain remodelling of PC, hydrolysis of LPC, and phospholipid metabolism pathways.
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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