Anti-Beta Tubulin TUBB Antibody Picoband® (monoclonal, 2E11)

Western blot analysis of Tubulin beta using anti-Tubulin beta antibody (M05613-4).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human HepG2 whole cell lysates,
Lane 3: human K562 whole cell lysates,
Lane 4: human Raji whole cell lysates,
Lane 5: rat brain tissue lysates,
Lane 6: rat lung tissue lysates,
Lane 6: mouse brain tissue lysates,
Lane 6: mouse lung tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-Tubulin beta antigen affinity purified monoclonal antibody (Catalog # M05613-4) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:10000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1001) with Tanon 5200 system. A specific band was detected for Tubulin beta at approximately 55 kDa. The expected band size for Tubulin beta is at 50 kDa.

IF analysis of Tubulin beta using anti-Tubulin beta antibody (M05613-4) and anti-FOSB antibody (A01569-1).
Tubulin beta was detected in immunocytochemical section of CACO-2 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL mouse anti-Tubulin beta Antibody (M05613-4) and rabbit anti-FOSB antibody (A01569-1) overnight at 4°C. DyLight®594 Conjugated Goat Anti-Mouse IgG (BA1141) and DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) were used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

IF analysis of Tubulin beta using anti-Tubulin beta antibody (M05613-4).
Tubulin beta was detected in immunocytochemical section of U2OS cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL mouse anti-Tubulin beta Antibody (M05613-4) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

IF analysis of Tubulin beta using anti-Tubulin beta antibody (M05613-4).
Tubulin beta was detected in immunocytochemical section of U2OS cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL mouse anti-Tubulin beta Antibody (M05613-4) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG (BA1126) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Flow Cytometry analysis of U937 cells using anti-Tubulin beta antibody M05613-4).
Overlay histogram showing U937 cells stained with M05613-4 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-Tubulin beta Antibody (M05613-4, 1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Flow Cytometry analysis of HEPA1-6 cells using anti-Tubulin beta antibody M05613-4).
Overlay histogram showing HEPA1-6 cells stained with M05613-4 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-Tubulin beta Antibody (M05613-4, 1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (BA1126, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Western blot analysis of Tubulin beta using anti-Tubulin beta antibody (M05613-4).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human Hela whole cell lysates,
Lane 2: human HepG2 whole cell lysates,
Lane 3: human K562 whole cell lysates,
Lane 4: human Raji whole cell lysates,
Lane 5: rat brain tissue lysates,
Lane 6: rat lung tissue lysates,
Lane 7: mouse brain tissue lysates,
Lane 8: mouse lung tissue lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-Tubulin beta antigen affinity purified monoclonal antibody (M05613-4) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-DyLight 647 Conjugated secondary antibody at a dilution of 1:2000 for 1.5 hour at RT. A specific band was detected for Tubulin beta at approximately 50 kDa. The expected band size for Tubulin beta is at 50 kDa.