Purification notesab76942 was affinity purified using an epitope specific to PTPN12 immobilized on solid support. Antibody concentration was determined by extinction coefficient: absorbance at 280 nm of 1.4 equals 1.0 mg of IgG.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human stomach carcinoma (left) and colon carcinoma (right) tissues labelling PTPN12 with ab76942 at 1/200 (1µg/ml). Detection: DAB.
Western blot - PTPN12 antibody (ab76942)
All lanes : Anti-PTPN12 antibody (ab76942) at 0.04 µg/ml
Lane 1 : HeLa whole cell lysate at 50 µg Lane 2 : HeLa whole cell lysate at 15 µg Lane 3 : HeLa whole cell lysate at 5 µg Lane 4 : mouse NIH3T3 whole cell lysate at 50 µg
Lane 1: ab76942, at 1 µg/mg, staining PTPN12 in HeLa whole cell lysate by Western blot of immunoprecipitate (1 mg of lysate for IP, 20% of IP loaded). Immunoprecipitation was performed by using ab76942 at 3µg/mg of lysate.
Lane 2: Control IgG.
Detection: Chemiluminescence with an exposure time of 3 seconds.
ICC/IF image of ab76942 stained A549 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 (ab76942, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 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.
References for Anti-PTPN12 antibody (ab76942)
This product has been referenced in:
Sztacho M et al. BAR Proteins PSTPIP1/2 Regulate Podosome Dynamics and the Resorption Activity of Osteoclasts. PLoS One11:e0164829 (2016).
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