Western blot analysis of Fluorescein Isothiocyanate (FITC) was performed by loading various amounts of a FITC-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 PVDF membrane using the G2 Fast Blotter (Product # 62288) and blocked with 5% milk in TBST for at least 1 hour at room temperature. FITC-BSA was detected at ~72kD using a FITC monoclonal antibody (Product # MIF2901) at a dilution of 1ug/ml in 5% milk in TBST overnight at 4C on a rocking platform, followed by a goat anti-mouse IgG-HRP secondary antibody (Product # 31430) at a dilution of 1:50,000 for at least 1 hour. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34076).
|Tested species reactivity||Chemical, Tag|
|Host / Isotype||Mouse / IgG1|
|Storage buffer||PBS, pH 7.4|
|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)||Assay Dependent|
|Immunoprecipitation (IP)||Assay Dependent|
|Radioimmune Assays (RIA)||Assay Dependent|
|Western Blot (WB)||1:1000|
* 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.
MIF2901 was formerly sold as a Seradyn product. This is a larger package size of MA5-14709.
Fluorescein isothiocyanate (FITC) is a derivative of fluorescein used in wide-ranging applications including flow cytometry. FITC is the original fluorescein molecule functionalized with an isothiocyanate reactive group (-N=C=S), replacing a hydrogen atom on the bottom ring of the structure. This derivative is reactive towards nucleophiles including amine and sulfhydryl groups on proteins.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.