[IF=2.74] Rui Feng. et al. The ameliorative effect of melatonin on LPS-induced Sertoli cells inflammatory and tight junctions damage via suppression of the TLR4/MyD88/NF-κB signaling pathway in newborn calf. Theriogenology. 2022 Feb;179:103  IF ;  Bovine.  

PubMed:34871925

[IF=2.33] Xiao, Tangli, et al. "Rapamycin promotes podocyte autophagy and ameliorates renal injury in diabetic mice." Molecular and Cellular Biochemistry (2014): 1-10.  IHC-P ;  Mouse.  

PubMed:24850187

[IF=2.299] Wang X et al. 3, 3′, 5-Triiodo-L-thyronine affects polarity proteins of bovine Sertoli cells via WT1/non-canonical Wnt signaling pathway. Theriogenology. 2020 Feb 21;148:8-17.  WB ;  Bovine.  

PubMed:32126395

[IF=2.299] Wang X et al. Influence of Wilms' tumor suppressor gene WT1 on bovine Sertoli cells polarity and tight junctions via non-canonical WNT signaling pathway. Theriogenology. 2019 Jul 8;138:84-93.  ICF ;  Cow.  

PubMed:31302435

[IF=1.723] Wang X et al. Wilms' tumour 1 (WT1) negatively regulates the expression of connexin 43 via a non-canonical Wnt signalling pathway in cultured bovine Sertoli cells. Reprod Fertil Dev. 2020 Feb 6.  WB&ICF ;  Bovine.  

PubMed:32023428

Sample:
Lane 1: Mouse Testis tissue lysates
Lane 2: Mouse Ovary tissue lysates
Lane 3: Mouse Liver tissue lysates
Lane 4: Human HL-60 cell lysates
Primary: Anti-WT1 (bs-6983R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 55 kDa
Observed band size: 50 kDa

Sample: TM4 Cell (Mouse) Lysate at 40 ug
Primary: Anti-Wilms Tumor Protein (bs-6983R) at 1/300 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 55 kD
Observed band size: 48 kD

Tissue/cell: human kidney 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-WT-1/Wilms Tumor Protein Polyclonal Antibody, Unconjugated(bs-6983R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(C-0010) staining

Paraformaldehyde-fixed, paraffin embedded (Mouse kidney); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Wilms Tumor Protein) Polyclonal Antibody, Unconjugated (bs-6983R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (Rat testis); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Wilms Tumor Protein) Polyclonal Antibody, Unconjugated (bs-6983R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (Mouse testis); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Wilms Tumor Protein) Polyclonal Antibody, Unconjugated (bs-6983R) at 1:400 overnight at 4°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (bs-0295G-FITC) for 90 minutes, and DAPI for nuclei staining.

Blank control:Molt-4.
Primary Antibody (green line): Rabbit Anti-Wilms Tumor Protein antibody (bs-6983R)
Dilution: 1μg /10^6 cells;
Isotype Control Antibody (orange line): Rabbit IgG .
Secondary Antibody : Goat anti-rabbit IgG-AF647
Dilution: 1μg /test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at-20℃. The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at 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.