Antibody Type: Recombinant Monoclonal

Application: ELISA, WB, IHC, IF, FC

Reactivity: Human

Rabbit IgG

9Q805

Human

1. Western Blot
-Positive WB detected in A375 whole cell lysate
-All lanes CD146 antibody at 0.6μg/ml
-Secondary: Goat polyclonal to rabbit IgG at 1/50000 dilution
-Predicted band size: 72 KDa
-Observed band size: 120 KDa
2. IHC image of TMAH-00170 diluted at 1:100 and staining in paraffin-embedded human breast 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 biotinylated secondary antibody and visualized using an HRP conjugated SP system.
3. Immunofluorescence staining of U251 cells with TMAH-00170 at 1:37,counter-stained with DAPI. The cells were fixed in 4% formaldehyde, permeabilized using 0.2% Triton X-100 and blocked in 10% normal Goat Serum. The cells were then incubated with the antibody overnight at 4°C.The secondary antibody was Alexa Fluor 488-congugated AffiniPure Goat Anti-Rabbit IgG (H+L).
4. Overlay histogram showing A375 cells stained with TMAH-00170 (red line) at 1:50. The cells were fixed with 70% Ethylalcohol (18h) and then permeabilized with 0.3% Triton X-100 for 2 min. The cells were then incubated in 1x PBS /10% normal goat serum to block non-specific protein-protein interactions followed by primary antibody for 1 h at 4°C.The secondary antibody used was FITC goat anti-rabbit IgG (H+L) at 1/200 dilution for 1 h at 4°C. Control antibody (green line) was used under the same conditions. Acquisition of >10,000 events was performed.

ELISA, WB, IHC, IF, FC

WB:1:500-1:5000; IHC:1:50-1:500; IF:1:30-1:200.

Monoclonal

Unconjugated

A synthetic peptide

Human

4162

Immunology