ROBO1

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

Activity
Measured by its binding ability in a functional ELISA.Immobilized Human ROBO1 at 2 μg/ml can bind Anti-ROBO1 recombinant antibody (CSB-RA896760MA1HU).The EC₅₀ is 0.7363-0.9105 ng/mL.

The purity of ROBO1 was greater than 90% as determined by SEC-HPLC

IHC image of CSB-RA896760MA1HU diluted at 1:30 and staining in paraffin-embedded human brain 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-Mouse IgG labeled by HRP and visualized using 0.05% DAB.

IHC image of CSB-RA896760MA1HU diluted at 1:30 and staining in paraffin-embedded human gastric 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-Mouse IgG labeled by HRP and visualized using 0.05% DAB.

Immunofluorescence staining of A549 cell with CSB-RA896760MA1HU at 1:20, counter-stained with DAPI. The cells were fixed in 4% formaldehyde and blocked in 10% normal Goat Serum. The cells were then incubated with the antibody overnight at 4C. The secondary antibody was FITC-conjugated AffiniPure Goat Anti-Mouse IgG(H+L).

Immunofluorescence staining of Hela cell with CSB-RA896760MA1HU at 1:20, counter-stained with DAPI. The cells were fixed in 4% formaldehyde and blocked in 10% normal Goat Serum. The cells were then incubated with the antibody overnight at 4C. The secondary antibody was FITC-conjugated AffiniPure Goat Anti-Mouse IgG(H+L).

Overlay Peak curve showing Hela cells surface stained with CSB-RA896760MA1HU (red line) at 1:50. Then 10% normal goat serum was Incubated to block non-specific protein-protein interactions followed by the antibody (1µg/1*10⁶cells) for 45 min at 4°C. The secondary antibody used was FITC-conjugated Goat Anti-Mouse IgG(H+L) at 1/200 dilution for 35 min at 4°C. Isotype control antibody (green line) was mouse IgG1 (1µg/1*10⁶cells) used under the same conditions. Acquisition of >10,025 events was performed.

Western Blot
Positive WB detected in: Hela whole cell lysate
All lanes: ROBO1 antibody at 3.2µg/ml
Secondary
Goat polyclonal to rabbit IgG at 1/50000 dilution
Predicted band size: 181, 182, 177, 176, 170 kDa
Observed band size: 181 kDa

Immunofluorescent analysis of Hela cells using CSB-PA896760LA01HU at dilution of 1:100 and Alexa Fluor 488-congugated AffiniPure Goat Anti-Rabbit IgG(H+L)