Western blot analysis of TGF beta-1 was performed by loading the indicated amounts of recombinant human active or latent TGF beta-1 protein, 5ul of serum samples from multiple sclerosis (MS) and leukemia patients, and 15ul of PageRuler Prestained Protein Ladder (Product # 26616) per well onto a Novex® 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane using the G2 Fast Blotter (Product # 62288), and blocked with StartingBlock (PBS) Blocking Buffer (Product # 37538) for 1 hour at room temperature. TGF beta-1 was detected at ~12kD (monomer) and ~24kD (homodimer) using a TGF beta-1 polyclonal antibody (Product # PA1-197) at a dilution of 1:250 in blocking buffer overnight at 4C on a rocking platform, followed by a goat anti-rabbit IgG-HRP secondary antibody (Product # 31460) at a dilution of 1:10,000 for at least 1hour at room temperature. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34080).
|Tested species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Full length human TGF beta-1 protein (aa 279-390)|
|Storage buffer||PBS with 1mg/ml BSA, 30% glycerol|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Western Blot (WB)||1:250-1:1000|
* 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.
By Western blot, PA1-197 detects both TGF beta-1 monomer and homodimer.
TGF beta-1 is a polypeptide member of the transforming growth factor beta superfamily of cytokines, found almost ubiquitously in tissues. Transforming growth factor (TGF)-b is stored in the extracellular matrix as a latent complex with its prodomain. Activation of TGF-b1 requires the binding of aV integrin to an RGD sequence in the prodomain and exertion of force on this domain, which is held in the extracellular matrix by latent TGF-b binding proteins. Latent forms are complexes of TGF-beta, an aminoterminal portion of the TGF-beta precursor, designated TGF-LAP (TGF-latency associated peptide), and a specific binding protein, known as LTBP.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.