Western blot analysis of human TGF beta-1 was performed by loading 10ul of human serum from patients with multiple sclerosis (MS), leukemia, or asthma (left panel) and 2ug and 1ug of recombinant human TGF beta-1 (right panel) protein per well on a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with StartingBlock (TBS) Blocking Buffer (Product # 37542) for at least 1 hour. The membrane was probed with a TGF beta-1 monoclonal antibody (Product # MA1-116) at a concentration of 5ug/ml overnight at 4C on a rocking platform, washed in TBS-0.1%Tween-20, and probed with a goat anti-mouse IgG-HRP secondary antibody (Product # 31430) at a dilution of 1:10,000 for at least 1 hour. Chemiluminescent detection was performed using SuperSignal West Dura (Product # 34075).
|Tested species reactivity||Human|
|Host / Isotype||Mouse / IgG1|
|Immunogen||Full length of human TGF beta-1 (Met1-Ser390) was fused with c-terminal polyhistidine tag|
|Tested Applications||Dilution *|
|Western Blot (WB)||1ug/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.
MA1-116 has successfully been used as a detection antibody in a sandwich ELISA with Product # MA1-169 used as a coating antibody.
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.