Western blot analysis of ECT was performed using whole cell lysates and tissue lysate of Caco-2 (Lane 1), Hep G2 (Lane 2) and Mouse heart (Lane 3). The blot was probed with Anti-ECT Rabbit polyclonal Antibody (Product # PA1-545, 2 µg/ml) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.4 µg/ml, 1:2500 dilution). A ~ 43 kDa band corresponding to ECT was observed across cell lines and tissue lysate tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody using iBind™ Flex Western Starter Kit (Product # SLF2000S). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
|Tested species reactivity||Human, Mouse, Non-human primate, Rat|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic Peptide: C(157) T K A H H S S Q E M S S E Y R E(172)|
|Storage buffer||tris with glycine, 1mg/ml BSA|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Western Blot (WB)||2-4 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
PA1-545 detects ethanolaminephosphate cytidyltransferase (ECT) protein in human, rat, mouse and non-human primate samples.
PA1-545 has successfully been used in Western blot procedures. By Western blot, this antibody detects an ~50 kDa protein representing ECT from C3H/10T1/2 cell lysates.
PA1-545 immunizing peptide corresponds to amino acid residues 157-172 from human ECT protein. This immunizing peptide (PEP-241) can be used for neutralization and control experiments.
ECT (ethanolaminephosphate cytidyltransferase) is an important regulatory enzyme in the pathway for phosphatidylethanolamine (PE) biosynthesis. PE functions in cell signalling, membrane fusion, platelet activation, and cell cycle progression. Specialized forms of PE exist as plasmologens, natural cannabinoid anadimade, and glycosylated PE. The ECT enzyme acts in the second of three catalysis steps to produce CDP-ethanolamine from phosphoethanolamine (P-Etn). Study of ECT could help determine the role of plasmologens in disease, specifically lipid disorders such as peroxisomal diseases as well as neuronal degeneration associated with Alzheimer's disease.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.