Western blot analysis was performed on 5ul Human Plasma (Lane 1).The blot was probed with PON1 Mouse Monoclonal Antibody (Product# LFMA0099, 4ug/ml) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 ug/ml, 1:4000 dilution). A 40 kDa band corresponding to PON1 was observed in Human Plasma tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX), 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 following blocking with 5% skimmed milk. Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
|Tested species reactivity||Human, Mouse, Rat|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Purified, His-tagged, recombinant, human PON1 protein expressed in E. coli.|
|Storage buffer||HEPES with 0.15M NaCl, 0.01% BSA, 50% glycerol|
|Contains||0.03% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Western Blot (WB)||2-5 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.
A suggested positive control for this product is HepG2 cells.
The paraoxonase (PON) gene-family in humans has three members, PON1, PON2, and PON3, aligned next to each other on the long arm of chromosome 7q21.3-22.1. PON1 of these is synthesized primarily in the liver and a portion is secreted into the plasma, where it is associated with high density lipoproteins (HDL). And It hydrolyzes the active metabolites of several other organophosphorus insecticides (e.g., chlorpyrifos oxon, diazoxon), as well as nerve agents such as sarin, soman and VX (lethal nerve agent). The PON1 polymorphism may be responsible for neurodegeneration and considered to be an independent risk factor for Parkinson's disease.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.