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Western blot analysis of L-Thyroxine was performed by loading 600ng, 300ng, 150ng and 75ng of a Thyroxine-BSA conjugate and 10ul 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 # MIT0501 (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||0.09% 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.
MIT0501 targets in ELISA, Western Blot, IP, and RIA applications and shows reactivity with Chemical samples.
MIT0501 was 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.