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

Western Blot
Positive WB detected in: HepG2 whole cell lysate(30μg), K562 whole cell lysate(30μg), COLO205 whole cell lysate(30μg), MCF7 whole cell lysate(30μg), Jurkat whole cell lysate(30μg), A549 whole cell lysate(30μg), PC-3 whole cell lysate(30μg), U251 whole cell lysate(30μg), Mouse brain tissue lysate(30μg)
All lanes:DLAT antibody at 1:1000
Secondary
Goat polyclonal to rabbit IgG at 1/40000 dilution
Predicted band size: 69 kDa
Observed band size: 69 kDa
Exposure time：2min

IHC image of CSB-RA283972A0HU diluted at 1:100 and staining in paraffin-embedded human kidney tissue performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a Goat anti-rabbit polymer IgG labeled by HRP and visualized using 0.05% DAB.

IHC image of CSB-RA283972A0HU diluted at 1:100 and staining in paraffin-embedded human liver cancer performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4°C overnight. The primary is detected by a Goat anti-rabbit polymer IgG labeled by HRP and visualized using 0.05% DAB.

Overlay Peak curve showing MCF-7 cells stained with CSB-RA283972A0HU (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 FITC-conjugated goat anti-rabbit IgG (H+L) at 1/200 dilution for 35min at 4℃.Control antibody (green line) was Rabbit IgG (1ug/1*10⁶cells) used under the same conditions. Acquisition of >10, 000 events was performed.

Western Blot
Positive WB detected in: HepG2 whole cell lysate
All lanes: DLAT antibody at 4μg/ml
Secondary
Goat polyclonal to rabbit IgG at 1/50000 dilution
Predicted band size: 69 kDa
Observed band size: 69 kDa

Immunoprecipitating DLAT in HepG2 whole cell lysate
Lane 1: Rabbit control IgG instead of CSB-PA006926LA01HU in HepG2 whole cell lysate. For western blotting, a HRP-conjugated Protein G antibody was used as the secondary antibody (1/2000)
Lane 2: CSB-PA006926LA01HU (6μg) + HepG2 whole cell lysate (1mg)
Lane 3: HepG2 whole cell lysate (20μg)