(Tris-Glycine gel) Discontinuous SDS-PAGE (reduced) with 5% enrichment gel and 15% separation gel.

Western Blot
Positive WB detected in: MCF-7 whole cell lysate(30μg), A-431 whole cell lysate(30μg), HEK293 whole cell lysate(30μg), colo205 whole cell lysate(30μg)
All lanes: METRNL antibody at 1:1000
Secondary
Goat polyclonal to human IgG at 1/40000 dilution
Predicted band size: 34 kDa
Observed band size: 34 kDa
Exposure time: 3min

Overlay Peak curve showing Hela cells stained with CSB-RA731035MA1HU (red line) at 1:100. The cells were fixed in 4% formaldehyde and permeated by 0.2% TritonX-100 for 10min. Then 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (1ug/1*10⁶cells) for 45min at 4℃. The secondary antibody used was Fluorescein (FITC) AffiniPure Goat Anti-Human IgG, Fcγ fragment specific at 1:200 dilution for 35 min at 4℃.Control antibody (green line) was human IgG1 (1ug/1*10⁶cells) used under the same conditions. Acquisition of >10,000 events was performed.

Western Blot
Positive WB detected in: Hela whole cell lysate, U87 whole cell lysate, K562 whole cell lysate, PC3 whole cell lysate, Mouse brain tissue, Rat kidney tissue, Rat brain tissue
All lanes: METRNL antibody at 1:2000
Secondary
Goat polyclonal to rabbit IgG at 1/50000 dilution
Predicted band size: 35, 26 KDa
Observed band size: 35 KDa
Exposure time：1min