Antibody Type: Mouse Monoclonal

Application: ELISA, WB, IF, FC, IP

Reactivity: Human, Mouse, Rat, Rabbit

IgG1

8H69

Human, Mouse, Rat, Rabbit

1. Western Blot
-Positive WB detected in: K562 whole cell lysate, Rabbit Skeletal Muscle tissue, Rabbit Kidney lysate
-All lanes ENO1 antibody at 1:10000
-Secondary: Goat polyclonal to mouse IgG at 1/10000 dilution
-Predicted band size: 47 KDa
-Observed band size: 47 KDa
-Exposure time: 1min
2. Western Blot
-Positive WB detected in: MCF-7 whole cell lysate, Hela whole cell lysate, Jurkat whole cell lysate, HepG2 whole cell lysate
-All lanes ENO1 antibody at 1:10000
-Secondary: Goat polyclonal to mouse  IgG at 1/10000 dilution
-Predicted band size: 47 KDa
-Observed band size: 47 KDa
-Exposure time: 10s
3. Western Blot
-Positive WB detected in: HepG2 whole cell lysate at 20µg, 10µg, 5µg, 2.5µg, 1.25µg, 0.625µg
-All lanes: ENO1 antibody at 1:5000
-Secondary: Goat polyclonal to Mouse IgG at 1/10000 dilution
-Predicted band size: 47 kDa
-Observed band size: 47 KDa
-Exposure time: 10s
4. Western Blot
-Positive WB detected in: MCF-7 whole cell lysate
-All lanes: ENO1 antibody at 1:5000, 1:10000, 1:20000, 1:40000, 1:80000, 1:160000, 1:320000, 1:640000
-Secondary: Goat polyclonal to Mouse IgG at 1/10000 dilution
-Predicted band size: 47 kDa
-Observed band size: 47 KDa
-Exposure time: 10s
5. Immunofluorescence staining of MCF-7 cells with TMAH-00386 at 1:270, counter-stained with DAPI. The cells were blocked in 10% normal Goat Serum and then incubated with the primary antibody overnight at 4°C. The secondary antibody was Alexa Fluor 488-congugated AffiniPure Goat Anti-Mouse IgG(H+L).
6. Immunofluorescence staining of Hela cells with TMAH-00386 at 1:270, counter-stained with DAPI. The cells were blocked in 10% normal Goat Serum and then incubated with the primary antibody overnight at 4°C. The secondary antibody was Alexa Fluor 488-congugated AffiniPure Goat Anti-Mouse IgG(H+L).
7. Overlay histogram showing MCF-7 cells stained with TMAH-00386 (red line) at 1:550. The cells were 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-mouse IgG(H+L) at 1/200 dilution for 1 h at 4°C. Isotype control antibody (green line) was used under the same conditions. Acquisition of >10,000 events was performed.
8. Overlay histogram showing Hela cells stained with TMAH-00386 (red line) at 1:550. The cells were 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-mouse IgG(H+L) at 1/200 dilution for 1 h at 4°C. Isotype control antibody (green line) was used under the same conditions. Acquisition of >10,000 events was performed.
9. Immunoprecipitating ENO1 in HepG2 whole cell lysate
-Lane 1: Mouse control IgG (1µg) instead of TMAH-00386 in HepG2 whole cell lysate.
For western blotting, a HRP-conjugated Protein G antibody was used as the secondary antibody (1/2000)
-Lane 2: TMAH-00386 (1µl) + HepG2 whole cell lysate (500µg)
-Lane 3: HepG2 whole cell lysate (10µg)

ELISA, WB, IF, FC, IP

Monoclonal

Unconjugated

Recombinant Protein: Human Alpha-enolase Protein (2-434AA)

Human

2023

Immunology