Antibody Type: Recombinant Mouse Monoclonal

Application: ELISA, WB, IHC, IF, FCM

Reactivity: Human

Mouse IgG2a

2H864

Human

1. The Binding Activity of CD147 with Anti-CD147 recombinant Antibody
Activity: Measured by its binding ability in a functional ELISA. Immobilized Human CD147 at 2 μg/ml can bind Anti-CD147 recombinant Antibody, the EC50 is 21.95-33.12 ng/ml.
2. Western Blot
-Positive WB detected in: HepG2 whole cell lysate, ntera2 whole cell lysate, A549 whole cell lysate, U251 whole cell lysate
-All lanes: CD147 antibody at 1:1000
-Secondary: Goat polyclonal to mouse IgG at 1/50000 dilution
-Predicted band size: 42, 29, 23, 19 KDa
-Observed band size: 35, 50-60 KDa
3. IHC image of TMAH-00171 diluted at 1:200 and staining in paraffin-embedded human testis 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 polymer IgG labeled by HRP and visualized using 0.05% DAB.
4. IHC image of TMAH-00171 diluted at 1:200 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-mouse polymer IgG labeled by HRP and visualized using 0.05% DAB.
5. IHC image of TMAH-00171 diluted at 1:200 and staining in paraffin-embedded human placenta 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 polymer IgG labeled by HRP and visualized using 0.05% DAB.
6. IHC image of TMAH-00171 diluted at 1:200 and staining in paraffin-embedded human stomach 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 polymer IgG labeled by HRP and visualized using 0.05% DAB.
7. Immunofluorescence staining of Hela cells with TMAH-00171 at 1:150, counter-stained with DAPI. The cells were fixed in 4% formaldehyde and blocked in 10% normal Goat Serum. The cells were incubated with the antibody overnight at 4°C. Nuclear DNA was labeled in blue with DAPI. The secondary antibody was FITC-conjugated AffiniPure Goat Anti-Mouse IgG (H+L).
8. Overlay Peak curve showing Hela cells stained with TMAH-00171 (red line) with 1 μg/well (10 μg/mL, 100 μL/well). Then 10% normal goat serum was Incubated to block non-specific protein-protein interactions followed by the antibody (1µg/1*10^6 cells) 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*10^6 cells) used under the same conditions.Acquisition of >10,000 events was performed.
9. Overlay Peak curve showing Jurkat cells stained with TMAH-00171 (red line) with 1 μg/well (10 μg/mL, 100 μL/well). Then 10% normal goat serum was Incubated to block non-specific protein-protein interactions followed by the antibody (1µg/1*10^6 cells) 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*10^6 cells) used under the same conditions.Acquisition of >10,000 events was performed.
10. Overlay Peak curve showing HepG2 cells surface stained with TMAH-00171 (red line) with 1 μg/well (10 μg/mL, 100 μL/well). Then 10% normal goat serum was Incubated to block non-specific protein-protein interactions followed by the antibody (1µg/1*10^6 cells) 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*10^6 cells) used under the same conditions.Acquisition of >10,000 events was performed.

ELISA, WB, IHC, IF, FCM

WB:1:500-1:2000; IHC:1:50-1:200; IF:1:50-1:200; FCM:1:50-1:200.

Monoclonal

Unconjugated

Recombinant Protein: Human CD147 Protein

Human

682

Cancer