Antibody Type: Rabbit Polyclonal

Application: WB,IHC-P,IHC-Fr,IF,FCM

Reactivity: Human,HPV16

IgG

Human,HPV16

1. 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 (TMAB-00887) 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 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).
2. 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 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (HPV16 E7) Polyclonal Antibody, Unconjugated (TMAB-00887-2) at 1:2000 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
3. 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 min; Blocking buffer (normal goat serum) at 37°C for 30 min; Antibody incubation with (HPV16 E7) Polyclonal Antibody, Unconjugated (TMAB-00887) at 1:2000 overnight at 4°C, followed by operating according to SP Kit (Rabbit) instructionsand DAB staining.
4. Tissue/cell: Human parotid tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer (0.01M, pH6.0), 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 (TMAB-00887) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody and DAb staining.
5. Blank control: Hela.
Primary Antibody (green line): Rabbit Anti-HPV16 E7 antibody (TMAB-00887)
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.
6. Sample:
Lane 1: Recombinant HPV16 E7 protein Bacterial lysate, DsbC & His
Primary: Anti-HPV16 E7 (TMAB-00887) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 11 kDa
Observed band size: 61 kDa

WB,IHC-P,IHC-Fr,IF,FCM

WB: 1:500-2000; IHC-P: 1:100-500; IHC-Fr: 1:100-500; IF: 1:100-500; FCM: 1ug/Test

Polyclonal

KLH conjugated synthetic peptide: human HPV16 E7

Human