For the best experience on the Abcam website please upgrade to a modern browser such as Google Chrome
Our Abpromise guarantee covers the use of ab15537 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use a concentration of 5 µg/ml.|
|WB||Use at an assay dependent concentration.
The immunogen for ab15537 is located close to C-terminus and therefore should be able to detect the proform at 50 kDa and mature form at 12.5 kDa under reducing condition.
|IHC-P||1/100. Incubate for 10 minutes at RT. Staining of formalin-fixed tissues requires boiling tissue sections in 10mM citrate buffer, pH 6.0 for 10 minutes followed by cooling at RT for 20 minutes. Ab15537 demonstrates cytoplasmic staining.|
|IP||Use at an assay dependent concentration.|
The blot was blocked with 5% milk for 1 hour at 21°C and incubated with the primary antibody for 12 hours at 4°C.
Detects a 50 kDa species, which is the presumed TGF-Beta 3 precursor, across all cell lysates and species tested including: Hu, AGMk Ms, Rat, and Hm. Higher molecular weight species are observed in all lysates, which could be glycosylated forms of the precursor. A fast migrating species (~14 kDa) was observed in U2OS cell lysate (Lane 2), which could be the cleaved, active form of TGF-Beta3.
ICC/IF image of ab15537 stained HepG2 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab15537 5µg/ml overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti- rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"