VCP Mouse Monoclonal Antibody

• Catalog Number: orb783397
• Category: Antibodies
• Description: VCP Mouse Monoclonal Antibody
• Target: VCP
• Clonality: Monoclonal
• Species/Host: Mouse
• Isotype: IgG1,Κ
• Conjugation: Unconjugated
• Reactivity: Human, Mouse, Rat
• Predicted Reactivity: Mouse, Rat
• Form/Appearance: Liquid
• Concentration: 1mg/ml
• Buffer/Preservatives: 0.01M TBS (pH7.4) with 1% rAlbumin, 0.02% Proclin300 and 50% Glycerol.
• Immunogen: Recombinant human VCP.
• UniProt ID: P55072
• MW: 97 kDa
• Tested applications: FC, ICC, IF, IHC-Fr, IHC-P, WB
• Dilution range: WB=1:500-2000, IHC-P=1:25, IHC-F=1:25, ICC/IF=1:25, IF=1:25, Flow-Cyt=1:25
• Antibody Type: Primary Antibody
• Clone Number: N5A9
• 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.
• Alternative names: valosin-containing protein; 15S Mg(2+) ATPase p97
• Note: For research use only

Blank control: K562. Primary Antibody (green line): Mouse Anti-VCP antibody (orb783397), dilution: 1:25, Isotype Control Antibody (blue line): Mouse IgG, Secondary Antibody: Goat anti-mouse IgG-AF488, dilution: 1:400, 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.

Hela 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 (VCP) monoclonal Antibody, Unconjugated (orb783397) 1:25, 90 minutes at 37°C, followed by a conjugated Goat Anti-Mouse IgG antibody at 37°C for 90 minutes, Dylight® 554 Phalloidin (red) was used to stain the cell Cytoplasmic actin.

Paraformaldehyde-fixed, paraffin embedded (human breast carcinoma sections), Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min, Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes, Blocking buffer (normal goat serum) at 37°C for 30 min, Antibody incubation with (VCP) Monoclonal Antibody, Unconjugated (orb783397) at 1:200 overnight at 4°C, followed by operating according to SP Kit (Mouse) instructionsand DAB staining.

Sample: Lane 1: Hela cell lysates, Lane 2: K562 cell lysates, Lane 3: A549 cell lysates, Lane 4: C6 cell lysates, Lane 5: U-87 MG cell lysates, Lane 6: NIH/3T3 cell lysates, Primary: Anti-VCP (orb783397) at 1/4000 dilution, Secondary: IRDye800CW Goat Anti-Mouse IgG at 1/20000 dilution, Predicted band size: 97 kD, Observed band size: 97 kD.