Anti-Tubulin beta Antibody Picoband®

The effect of NAM on MAPK and NF-κB signaling pathway in lung. a – g Lung tissues from different experimental groups were obtained 12 h after LPS administration and total protein extracted. The phosphorylation levels of ERK, JNK, P38, AKT, IκBα and P65 were determined via western blot and quantitated via densitometry using β-tubulin as an internal control. The values are presented as the mean ± SD (* p < 0.05, ** p < 0.001, *** p < 0.001 and **** p < 0.0001)
Index in PubMed under a CC BY license. PMID: 34544355

Western blot analysis of Tubulin beta using anti-Tubulin beta antibody (A05397-1).
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 CACO-2 whole cell lysates,
Lane 5: human SH-SY5Y whole cell lysates,
Lane 6: human SiHa whole cell lysates,
Lane 7: human MCF-7 whole cell lysates,
Lane 8: human A549 whole cell lysates,
Lane 9: rat brain tissue lysates,
Lane 10: mouse brain 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 rabbit anti-Tubulin beta antigen affinity purified polyclonal antibody (Catalog # A05397-1) 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-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Tubulin beta at approximately 50 kDa. The expected band size for Tubulin beta is at 50 kDa.

IHC analysis of Tubulin beta using anti-Tubulin beta antibody (A05397-1).
Tubulin beta was detected in a paraffin-embedded section of human colon cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-Tubulin beta Antibody (A05397-1) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of Tubulin beta using anti-Tubulin beta antibody (A05397-1).
Tubulin beta was detected in a paraffin-embedded section of human testicular cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-Tubulin beta Antibody (A05397-1) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IF analysis of Tubulin beta using anti-Tubulin beta antibody (A05397-1).
Tubulin beta was detected in an immunocytochemical section of A549 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 5 μg/mL rabbit anti-Tubulin beta Antibody (A05397-1) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:500 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 HepG2 cells using anti-Tubulin beta antibody (A05397-1).
Overlay histogram showing HepG2 cells stained with A05397-1 (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 rabbit anti-Tubulin beta Antibody (A05397-1,1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit 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.