bs-8533R [Primary Antibody]
Rabbit  Anti-Vimentin Polyclonal Antibody
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Host: Rabbit

Target Protein: Vimentin

IR: Immunogen Range:371-466/466

Clonality: Polyclonal

Isotype: IgG

Entrez Gene: 7431

Swiss Prot: P08670

Source: KLH conjugated synthetic peptide derived from human Vimentin:371-466/466 

Purification: affinity purified by Protein A

Storage: 0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol. Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.

Background: This gene encodes a member of the intermediate filament family. Intermediate filamentents, along with microtubules and actin microfilaments, make up the cytoskeleton. The protein encoded by this gene is responsible for maintaining cell shape, integrity of the cytoplasm, and stabilizing cytoskeletal interactions. It is also involved in the immune response, and controls the transport of low-density lipoprotein (LDL)-derived cholesterol from a lysosome to the site of esterification. It functions as an organizer of a number of critical proteins involved in attachment, migration, and cell signaling. Mutations in this gene causes a dominant, pulverulent cataract.[provided by RefSeq, Jun 2009]

Size: 50ul

Concentration: 1mg/ml

Applications: WB=1:500-2000,IHC-F=1:100-500,Flow-Cyt=1μg/Test,ICC=1:100,IF=1:100-500,ELISA=1:5000-10000

Cross Reactive Species: Human,Mouse,Rabbit (predicted: Rat,Pig,Cow,Dog,Horse)

For research use only. Not intended for diagnostic or therapeutic use.

VALIDATION IMAGES
Sample: A549(Human) Cell Lysate at 30 ug Jurkat(Human) Cell Lysate at 30 ug Primary: Anti-Vimentin   (bs-8533R) 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 (bs-8533R) at 1/1000 dilution Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution Predicted band size: 51 kD Observed band size: 57 kD
Protein: lung(rabbit) lysate at 40ug; Primary: rabbit Anti-Vimentin (bs-8533R) at 1:300; Secondary: HRP conjugated Goat-Anti-rabbit IgG(bs-0295G-HRP) at 1: 5000; Predicted band size: 51 kD Observed band size: 51 kD
Sample: A549(Human) Cell Lysate at 30 ug HUVEC(Human) Cell Lysate at 30 ug U251(Human) Cell Lysate at 30 ug Primary: Anti-Vimentin (bs-8533R) at 1/1000 dilution Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution Predicted band size: 53 kD Observed band size: 53 kD
Sample: U-87MG (Human) Cell Lysate at 30 ug NIH/3T3 (Mouse) Cell Lysate at 30 ug Primary: Anti-Vimentin (bs-8533R) at 1/1000 dilution Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution Predicted band size: 51 kD Observed band size: 53 kD
Tissue/cell: U-87MG 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-8533R)antibody (bs-0295G-FITC) at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.
Tissue/cell: endothelial cells of umbilical artery;4% Paraformaldehyde-fixed; Blocking buffer (normal goat serum,C-0005) at 37℃ for 20 min; Incubation: Anti-Vimentin Polyclonal Antibody, Alexa Fluor 488 conjugated(bs-8533R-A488) 1:100, 60 minutes at 37℃. DAPI(5ug/ml,blue,C-0033) 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, C-0005) at 37°C for 20 min; Antibody incubation with (Vimentin) polyclonal Antibody, Unconjugated (bs-8533R) 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.
Tissue/cell: 293T 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-8533R) 1:200, 2 hours at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody (bs-0295G-FITC) at 37°C for 90 minutes, DAPI (5ug/ml, blue, C-0033) 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, C-0005) at 37°C for 20 min; Antibody incubation with (Vimentin) Polyclonal Antibody, Unconjugated (bs-8533R) 1:200, 2 hours at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody (bs-0295G-FITC) at 37°C for 90 minutes, DAPI (5ug/ml, blue, C-0033) was used to stain the cell nuclei.
Blank control:A549. Primary Antibody (green line): Rabbit Anti-Vimentin antibody (bs-8533R) Dilution: 1μg /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 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.
 
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