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[IF=5.201] Juan-Juan Zhu. et al. VAMP3 and SNAP23 as Potential Targets for Preventing the Disturbed Flow-Accelerated Thrombus Formation. Front Cell Dev Biol. 2020; 8: 576826  WB,IF,IHC ;  Human.  

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Sample: Hela(Human) Cell Lysate at 30 ug Hela KO Vimentin (Human) Cell Lysate at 30 ug Primary: Anti- Vimentin (bsm-33170M) at 1/1000 dilution Secondary: IRDye800CW Goat Anti-Mouse IgG at 1/20000 dilution Predicted band size: 53 kD Observed band size: 58 kD

25 ug total protein per lane of various lysates (see on figure) probed with Vimentin monoclonal antibody, unconjugated (bsm-33170M) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.

Paraformaldehyde-fixed, paraffin embedded Uterine body cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Vimentin Monoclonal Antibody, Unconjugated(bsm-33170M) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Mouse, sp-0024) and DAB (C-0010) staining.

Paraformaldehyde-fixed, paraffin embedded Human Uterus; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Vimentin Monoclonal Antibody, Unconjugated(bsm-33170M) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Mouse, sp-0024) and DAB (C-0010) staining.

Paraformaldehyde-fixed, paraffin embedded Human Kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Vimentin Monoclonal Antibody, Unconjugated(bsm-33170M) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Mouse, sp-0024) and DAB (C-0010) staining.

Paraformaldehyde-fixed, paraffin embedded Human Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Vimentin Monoclonal Antibody, Unconjugated(bsm-33170M) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Mouse, sp-0024) and DAB (C-0010) staining.

Paraformaldehyde-fixed, paraffin embedded Human Tonsil; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Vimentin Monoclonal Antibody, Unconjugated(bsm-33170M) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Mouse, sp-0024) and DAB (C-0010) staining.

Paraformaldehyde-fixed, paraffin embedded Human Cervical Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Vimentin Monoclonal Antibody, Unconjugated(bsm-33170M) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Mouse, sp-0024) and DAB (C-0010) staining.

Hela 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 (Vimentin) monoclonal Antibody, Unconjugated (bsm-33170M) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Mouse IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.

Blank control:Hela. Primary Antibody (green line): Mouse Anti-Vimentin antibody (bsm-33170M) Dilution: 1ug/Test; Secondary Antibody : Goat anti-Mouse IgG-FITC Dilution: 0.5ug/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 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.

4% Paraformaldehyde-fixed Hela (left) cell and Vimentin knockout Hela(right) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (Vimentin) monoclonal Antibody, unconjugated (bsm-33170M) 1:100, 90 min at 37°C; followed by conjugated Goat Anti-Mouse IgG antibody (green, bs-60296G-FITC) at 37°C for 90 min, DAPI (blue, C02-04002) was used to stain the cell nuclei.