Western blot analysis of L-Thyroxine was performed by loading 600ng, 300ng, 150ng and 75ng of a Thyroxine-BSA conjugate and 10µl PageRuler Plus Prestained Protein Ladder (Product # 26619) per well onto a 4-20% Tris-Glycine polyacrylamide gel. Proteins were transferred to a nitrocellulose membrane using the G2 Fast Blotter (Product # 62288) and blocked with 5% milk in TBST for at least 1 hour at room temperature. L- Thyroxine was detected at ~62kD using a L-Thyroxine monoclonal antibody antibody (Product # MA5-14716 (T4YCH clone8)) at a dilution of 1µg/ml in 5% milk in TBST overnight at 4C on a rocking platform, followed by a goat anti-mouse IgG-HRP Superclonal secondary antibody (Product # A28177) at a dilution of 1:5000 for at least 1 hour. Chemiluminescent detection will be performed using SuperSignal West Dura (Product # 34078).
|Tested species reactivity||Chemical|
|Host / Isotype||Mouse / IgG2b|
|Storage buffer||PBS with 1mg/ml BSA, 30% glycerol|
|Contains||0.05% sodium azide|
|Storage Conditions||Maintain refrigerated at 2-8°C for up to 6 months. For long term storage store at -20°C|
|Tested Applications||Dilution *|
|ELISA (ELISA)||1 µg/ml|
|Immunoprecipitation (IP)||Assay Dependent|
|Radioimmune Assays (RIA)||Assay Dependent|
|Western Blot (WB)||1 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.
MA5-14716 targets L-Thyroxine (T4) in ELISA, Western Blot, IP, and RIA applications and shows reactivity with Chemical samples.
Product MA514716 is a smaller package size of MIT0501 (formerly sold as a Seradyn product).
Levothyroxine, also L-thyroxine, synthetic T4, or 3,5,3',5'-tetraiodo-L-thyronine, is a synthetic form of thyroxine (thyroid hormone), used as a hormone replacement for patients with thyroid problems. The natural hormone is chemically in the chiral L-form, as is the pharmaceutical agent. Dextrothyroxine (D-thyroxine) briefly saw research as an anticholesterol agent but was pulled due to cardiac side-effects.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.