VALIDATION IMAGES
Image submitted by One World Lab validation program. U138 cells were stained with bs-0756R Rabbit Anti-Vimentin Polyclonal Antibody at 1:100 in PBS and incubated for one hour at room temperature, followed by secondary antibody incubation, DAPI staining and detection.
Formalin-fixed and paraffin embedded mouse kidney tissue labeled Anti-Vimentin Polyclonal Antibody, Unconjugated (bs-0756R) at 1:200, followed by conjugation to the secondary antibody and DAB staining
Formalin-fixed and paraffin embedded mouse mesenchymal stem cells labeled Anti-Vimentin Polyclonal Antibody, Unconjugated (bs-0756R) at 1:300, followed by conjugation to the secondary antibody and DAB staining
Human Jurkat cells probed with Vimentin Polyclonal Antibody, Unconjugated (bs-0756R) at 0.2ug for 30 minutes followed by incubation with a PE Conjugated secondary (green) for 30 minutes compared to control cells (blue), secondary only (light blue) and isotype control (orange).
This image was kindly submitted by Preethi Vijayaraj from UCLA; 4% PFA fixed cells, permeabilized with 0.1% Triton/TBS for 10 minutes, blocked with proteinblock (DAKO) for 30 minutes, stained with 1:100 primary antibody for 1 hour at room temperature followed by 1:200 AlexaFluor 488 secondary antibody for 45 minutes
Image provided by One World Lab validation program. U138 cells were stained with bs-0756R Rabbit Anti-Vimentin Polyclonal Antibody at 1:100 dilution.
20 μg proteins of cell lysates from U138, HeLa and A431 were separated by SDS-PAGE and transferred to PVDF membrane for western blotting analysis antibodies against protein VIM at dilution of 1:1000 with exposure time of 10 seconds.
Mouse spleen lysates probed with Anti-Vimentin Polyclonal Antibody, Unconjugated (bs-0756R) at 1:300 in 4˚C. Followed by conjugation to secondary antibody (bs-0295G-HRP) at 1:5000 90min in RT
Lane 1: Hela cell lysates; Lane 2: Hela KO Vimentin cell lysates probed with Vimentin Polyclonal Antibody, Unconjugated (bs-0756R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.
HeLa cells were stained with Vimentin Polyclonal Antibody, Unconjugated(bs-0756R) at 1:50 in PBS and incubated for two hours at 37°C followed by Goat Anti-Rabbit IgG (H+L) FITC conjugated secondary antibody. DAPI staining of the nucleus was done and then detected.
U-2 OS cells were stained with Vimentin Polyclonal Antibody, Unconjugated(bs-0756R) at 1:50 in PBS and incubated for two hours at 37°C followed by Goat Anti-Rabbit IgG (H+L) FITC conjugated secondary antibody. DAPI staining of the nucleus was done and then detected.
U251 cells were stained with Vimentin Polyclonal Antibody, Unconjugated(bs-0756R) at 1:50 in PBS and incubated for two hours at 37°C followed by Goat Anti-Rabbit IgG (H+L) FITC conjugated secondary antibody. DAPI staining of the nucleus was done and then detected.
Tissue/cell: HepG2 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) Polyclonal Antibody, Unconjugated (bs-0756R) 1:50, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody (bs-0295G-FITC) at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.
A549 cells were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at -20℃, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with Vimentin Polyclonal Antibody(bs-0756R)at 1:100 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).
Lane 1: Human Hela cell lysates; Lane 2: Mouse NIH/3T3 cell lysates; Lane 3: Human U251 cell lysates; Lane 4: Human SH-SY5Y cell lysates; Lane 5: Human A549 cell lysates probed with Vimentin Polyclonal Antibody, Unconjugated (bs-0756R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C
Paraformaldehyde-fixed, paraffin embedded (rat colon); 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 (Vimentin) Polyclonal Antibody, Unconjugated (bs-0756R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat ovary); 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 (Vimentin) Polyclonal Antibody, Unconjugated (bs-0756R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (mouse ovary); 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 (Vimentin) Polyclonal Antibody, Unconjugated (bs-0756R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (mouse brain); 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 (Vimentin) Polyclonal Antibody, Unconjugated (bs-0756R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat brain); 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 (Vimentin) Polyclonal Antibody, Unconjugated (bs-0756R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat 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 (Vimentin) Polyclonal Antibody, Unconjugated (bs-0756R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB 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 (Vimentin) Polyclonal Antibody, Unconjugated (bs-0756R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human lung carcinoma); 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 (Vimentin) Polyclonal Antibody, Unconjugated (bs-0756R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human colon carcinoma); 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 (Vimentin) Polyclonal Antibody, Unconjugated (bs-0756R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human breast carcinoma); 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 (Vimentin) Polyclonal Antibody, Unconjugated (bs-0756R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human 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 (Vimentin) Polyclonal Antibody, Unconjugated (bs-0756R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human tonsil ); 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 (Vimentin) Polyclonal Antibody, Unconjugated (bs-0756R) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded Mouse Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Vimentin Polyclonal Antibody, Unconjugated (bs-0756R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Rat Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Vimentin Polyclonal Antibody, Unconjugated (bs-0756R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) 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 Polyclonal Antibody, Unconjugated (bs-0756R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) 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 Polyclonal Antibody, Unconjugated (bs-0756R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Mouse Kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Vimentin Polyclonal Antibody, Unconjugated (bs-0756R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Rat Kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Vimentin Polyclonal Antibody, Unconjugated (bs-0756R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Small Intestine; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Vimentin Polyclonal Antibody, Unconjugated (bs-0756R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) 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 Polyclonal Antibody, Unconjugated (bs-0756R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) 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 Polyclonal Antibody, Unconjugated (bs-0756R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) 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 Polyclonal Antibody, Unconjugated (bs-0756R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Breast Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Vimentin Polyclonal Antibody, Unconjugated (bs-0756R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.
Paraformaldehyde-fixed, paraffin embedded Human Endometrium Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with Vimentin Polyclonal Antibody, Unconjugated (bs-0756R) at 1:200 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.
25 ug total protein per lane of various lysates (see on figure) probed with Vimentin polyclonal antibody, unconjugated (bs-0756R) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.