Vimentin Rabbit Polyclonal Antibody

• Catalog Number: orb158714
• Category: Antibodies
• Description: Vimentin Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, ICC, IF, IHC-Fr, IHC-P, WB
• Predicted Reactivity: Bovine, Canine, Equine, Porcine, Rabbit
• Reactivity: Human, Mouse, Rat
• CAS Number: Vimentin
• Isotype: IgG
• Immunogen: KLH conjugated synthetic peptide derived from human Vimentin (371-466/466aa)
• Concentration: 1mg/ml
• Dilution range: WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, ICC/IF=1:100, IF=1:100-500, Flow-Cyt=1μg/Test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 51 kDa
• Target: VIM
• UniProt ID: P08670
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Alternative names: VIM; FLJ36605; OTTHUMP00000019224; VIM; VIME_HUMAN
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

References

Wang, Wei et al. Anti-cancer effect of Aquaporin 5 silencing in colorectal cancer cells in association with inhibition of Wnt/β-catenin pathway Cytotechnology, 70, 615-624 (2018)

PubMed: 29455395

Blank control: A549. Primary Antibody (green line): Rabbit Anti-Vimentin antibody (orb158714), dilution: 1 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody: Goat anti-rabbit IgG-AF488, dilution: 1 µg/Test. Protocol, The cells were fixed with 4% PFA (10 min at room temperature) and then permeabilized with 90% ice-cold methanol for 20 min at -20°C. 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 20000 events was performed.

Sample: A549 (Human) Cell Lysate at 30 ug, Jurkat (Human) Cell Lysate at 30 ug, Primary: Anti-Vimentin (orb158714) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 51 kD, Observed band size: 53 kD.

Sample: Hela KO Vimentin (Human) Cell Lysate at 30 ug, Hela (Human) Cell Lysate at 30 ug, Primary: Anti-Vimentin (orb158714) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 51 kD, Observed band size: 57 kD.

Tissue/Cell: 293T cell, 4% Paraformaldehyde-fixed, Triton X-100 at room temperature for 20 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Antibody incubation with (Vimentin) Polyclonal Antibody, Unconjugated (orb158714) 1:200, 2 hours at 37°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (orb868805) at 37°C for 90 minutes, DAPI (5 ug/ml, blue) was used to stain the cell nuclei.

Tissue/Cell: endothelial cells of umbilical artery, 4% Paraformaldehyde-fixed, Blocking buffer (normal goat serum) at 37°C for 20 min, Incubation: Anti-Vimentin Polyclonal Antibody, Alexa Fluor 488 conjugated 1:100, 60 minutes at 37°C. DAPI (5 ug/ml, blue) was used to stain the cell nuclei.

Tissue/Cell: FHC cell, 4% Paraformaldehyde-fixed, Triton X-100 at room temperature for 20 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Antibody incubation with (Vimentin) Polyclonal Antibody, Unconjugated (orb158714) 1:200, 2 hours at 37°C, followed by a conjugated Goat Anti-Rabbit IgG antibody (orb868805) at 37°C for 90 minutes, DAPI (5 ug/ml, blue) was used to stain the cell nuclei.

Tissue/Cell: U-87MG cell, 4% Paraformaldehyde-fixed, Triton X-100 at room temperature for 20 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Antibody incubation with (Vimentin) polyclonal Antibody, Unconjugated (orb158714) 1:100, 90 minutes at 37°C, followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.

Tissue/Cell: U-87MG cell, 4% Paraformaldehyde-fixed, Triton X-100 at room temperature for 20 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Antibody incubation with (Vimentin) Polyclonal Antibody, Unconjugated (orb158714) antibody (orb868805) at 37°C for 90 minutes, DAPI (blue) was used to stain the cell nuclei.