[IF=3.503] Ying Yang. et al. Integrated chemical profiling, network pharmacology and pharmacological evaluation to explore the potential mechanism of Xinbao pill against myocardial ischaemia–reperfusion injury. Pharm Biol. 2022;60(1):255-273  WB ;  Rat.  

PubMed:35148221

[IF=3.241] Su H et al. cis 9, trans 11, but not trans 10, cis 12 CLA isomer, impairs intestinal epithelial barrier function in IPEC-J2 cells and mice through activation of GPR120-[Ca2+]i and the MLCK signaling pathway. Food Funct. 2020 Apr 30;11(4):3657-3667.  WB ;  pig.  

PubMed:32296804

[IF=3.23] Chen, Jiahuan, et al. "Isolation and characterization of porcine amniotic fluid-derived multipotent stem cells." PLoS One 6.5 (2011): e19964.  IF(ICC) ;  Pig.  

PubMed:21625520

[IF=2.58] Jiang, Xiao-Hua, et al. "Effects of hepatitis C virus core protein and nonstructural protein 4B on the Wnt/β-catenin pathway." BMC microbiology 17.1 (2017): 124.  WB ;  Human.  

PubMed:28545480

Sample:
Lane 1: Cerebellum (Rat) Lysate at 40 ug
Lane 2: Cerebellum (Mouse) Lysate at 40 ug
Lane 3: Cerebrum (Mouse) Lysate at 40 ug
Lane 4: Cerebrum (Rat) Lysate at 40 ug
Lane 5: Ovary (Mouse) Lysate at 40 ug
Lane 6: Heart (Mouse) Lysate at 40 ug
Lane 7: Heart (Rat) Lysate at 40 ug
Primary: Anti-beta Tubulin (bs-0210R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 50 kD
Observed band size: 50 kD

Tissue/cell: rat brain tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min;
Incubation: Anti-Tubulin-beta Polyclonal Antibody, Unconjugated(bs-0210R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining

Tissue/cell:Sh-sy5y cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (beta Tubulin) polyclonal Antibody, Unconjugated (bs-0210R) 1:100, 90 minutes at 37°C; followed by a FITC conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.

SH-SY5Y cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (beta Tubulin) polyclonal Antibody, Unconjugated (bs-0210R) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.

Blank control: RSC96(blue), the cells were fixed with 2% paraformaldehyde (10 min) and then permeabilized with ice-cold 90% methanol for 30 min on ice..
Isotype Control Antibody: Rabbit IgG(orange) ; Secondary Antibody: Goat anti-rabbit IgG-FITC(white blue), Dilution: 1:100 in 1 X PBS containing 0.5% BSA ; Primary Antibody Dilution: 1μl in 100 μL1X PBS containing 0.5% BSA(green).

Blank control:HL-60.
Primary Antibody (green line): Rabbit Anti-beta Tubulin antibody (bs-0210R)
Dilution: 2μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-AF488
Dilution: 1μg /test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 0.1%PBST for 20 min at room temperature. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.