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Western blot analysis of Phosphorylated Progesterone Receptor (upper panel) and total Progesterone Receptor (lower panel) was performed by loading 20ug of nuclear (N) or cytoplasmic (C) T47D cell lysates, untreated (-) or stimulated (+) with 100 nm promegestone (R5020) for 1 hour 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/TBST for at least 1 hour at room temperature. Phosphorylated Progesterone Receptor was detected using a Phospho-Progesterone Receptor (Ser190) mouse monoclonal antibody (upper panel, Product # 37-8200) at a dilution of 1:1000 and Total Progesterone Receptor was detected using a Progesterone Receptor mouse monoclonal antibody (lower panel, MA1-12626) at a concentration of 1ug/ml in blocking buffer overnight at 4°C on a rocking platform, followed by a Superclonal goat anti-Mouse IgG-HRP secondary antibody (Product # A28177) at a dilution of 1:2,000 for at least 1 hour at room temperature. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34078) and the myECL Imager (Product #62236).
|Tested species reactivity||Mouse|
|Host / Isotype||Goat / IgG|
|Immunogen||Recombinant full-length protein|
|Purification||Gravity column chromatography|
|Storage buffer||PBS, HRP Stabilizer|
|Storage Conditions||4° C, store in dark, DO NOT FREEZE!|
|Antibody Form||Whole Antibody|
|Tested Applications||Dilution *|
|ELISA (ELISA)||0.05-1 µg/ml|
|Flow Cytometry (Flow)||1-5 µg/ml|
|Western Blot (WB)||0.05-1 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.
The sensitivity and specificity of each lot is confirmed using ELISA.
Minimal cross-reactivity with rabbit, rat, human, bovine, guinea pig and donkey IgG is observed.
Superclonal™ secondary antibodies are developed to be highly specific for both heavy and light chains of IgG. Utilizing the power of recombinant technology and multiple phenotypic screens, Superclonal™ antibodies are selected for enhanced specificity to the target species IgG. These recombinant secondary antibodies show similar performance as traditional polyclonal secondary antibodies in common immunoassay applications, with minimal cross-reactivity to other commonly-used species. In addition, their recombinant animal-free-origin helps ensures lot-to-lot consistency, reducing the need to optimize each lot in relevant assays.
Superclonal™ secondary antibodies are designed to allow scientists to achieve the accurate results they need, without some of the concerns that can impact traditional polyclonal secondary antibodies.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.