HPV16 E7 Rabbit Polyclonal Antibody

• Catalog Number: orb184006
• Category: Antibodies
• Description: HPV16 E7 Rabbit Polyclonal Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: FC, IF, IHC-Fr, IHC-P, WB
• Reactivity: Human
• Isotype: IgG
• Immunogen: KLH conjugated synthetic peptide derived from human HPV16 E7 (21-98/98aa)
• Antibody Type: Primary Antibody
• Concentration: 1mg/ml
• Dilution range: WB=1:500-2000, IHC-P=1:100-500, IHC-F=1:100-500, IF=1:100-500, Flow-Cyt=1ug/Test
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• MW: 11 kDa
• Target: L2
• 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: E7; HPV16 E7 protein; Human Papilloma Virus; Human
• Note: For research use only
• Expiration Date: 12 months from date of receipt.

Black line: Positive blank control (Hela), Negative blank control (RSC96), Green line: Primary Antibody (Rabbit Anti-HPV16 E7 antibody), Orange line: Isotype Control Antibody (Rabbit IgG). Blue line: Secondary Antibody (Goat anti-rabbit IgG-AF647), Hela (Positive) and RSC96 (Negative control) cells (black) were fixed with 4% PFA for 10 min at room temperature, permeabilized with 90% ice-cold methanol for 20 min at -20°C, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with HPV16 E7 Antibody (orb184006) at 1:50 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2% BSA in PBS, followed by secondary antibody (blue) incubation for 40 min at room temperature. Acquisitions of 20000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).

Blank control: Hela. Primary Antibody (green line): Rabbit Anti-HPV16 E7 antibody (orb184006), dilution: 1 µg/10^6 cells, Isotype Control Antibody (orange line): Rabbit IgG. Secondary Antibody: Goat anti-rabbit IgG-PE, 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 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.

Paraformaldehyde-fixed, paraffin embedded (human cervical carcinoma), 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 (HPV16 E7) Polyclonal Antibody, Unconjugated (orb184006) at 1:2000 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Paraformaldehyde-fixed, paraffin embedded (human cervical carcinoma), 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 (HPV16 E7) Polyclonal Antibody, Unconjugated at 1:2000 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.

Sample: Lane 1: Recombinant HPV16 E7 protein Bacterial lysate, DsbC & His, Primary: Anti-HPV16 E7 (orb184006) at 1/1000 dilution, Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution, Predicted band size: 11 kDa, Observed band size: 61 kDa.

Tissue/Cell: Human parotid tissue, 4% Paraformaldehyde-fixed and paraffin-embedded, Antigen retrieval: citrate buffer (0.01M, pH 6.0), Boiling bathing for 15 min, Block endogenous peroxidase by 3% Hydrogen peroxide for 30 min, Blocking buffer (normal goat serum) at 37°C for 20 min, Incubation: Anti-HPV16 E7 Polyclonal Antibody, Unconjugated (orb184006) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAB staining.