Sample:
Cerebrum (Mouse) Lysate at 40 ug
Primary: Anti-PARK7 (SL1306R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 20 kD
Observed band size: 20 kD
Sample:
Lane1:Brain(Rat) lysate at 30ug;
Lane2:Brain(Rat) lysate at 30ug;
Primary: Anti-CAP1/PARK7 (SL1306R) at 1:200 dilution;
Secondary: Alkaline phosphatase conjugated Goat Anti-Rabbit IgG(SL0295G-AP) at 1: 3000 dilution;
Predicted band size : 20kD
Observed band size : 20kD
Sample:
Lane 1: Testis (Mouse) Lysate at 40 ug
Lane 2: Liver (Mouse) Lysate at 40 ug
Lane 3: Cerebrum (Rat) Lysate at 40 ug
Lane 4: Thyroid gland Rat) Lysate at 40 ug
Lane 5: Kidney (Rat) Lysate at 40 ug
Lane 6: Liver (Rat) Lysate at 40 ug
Lane 7: Hela (Human) Cell Lysate at 30 ug
Lane 8: U937 (Human) Cell Lysate at 30 ug
Lane 9: K562 (Human) Cell Lysate at 30 ug
Lane 10: HL60 (Human) Cell Lysate at 30 ug
Primary:
Anti-PARK7/DJ1 (SL1306R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 22 kD
Observed band size: 22 kD
Sample:
Jurkat(Human) Cell Lysate at 30 ug
Hela(Human) Cell Lysate at 30 ug
Primary: Anti-PARK7 (SL1306R) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution
Predicted band size: 20 kD
Observed band size: 20 kD
Paraformaldehyde-fixed, paraffin embedded (rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (PARK7) Polyclonal Antibody, Unconjugated (SL1306R ) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human colon carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (PARK7) Polyclonal Antibody, Unconjugated (SL1306R ) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining.
Tissue/cell: rat kidney tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,SLC0005) at 37℃ for 20 min;
Incubation: Anti-CAP1/PARK7 Polyclonal Antibody, Unconjugated(SL1306R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(SLC0010) staining
Tissue/cell: rat kidney tissue; 4% Paraformaldehyde-fixed and paraffin-embedded;
Antigen retrieval: citrate buffer ( 0.01M, pH 6.0 ), Boiling bathing for 15min; Block endogenous peroxidase by 3% Hydrogen peroxide for 30min; Blocking buffer (normal goat serum,SLC0005) at 37℃ for 20 min;
Incubation: Anti-CAP1/PARK7 Polyclonal Antibody, Unconjugated(SL1306R) 1:200, overnight at 4°C, followed by conjugation to the secondary antibody(SP-0023) and DAB(SLC0010) staining
Blank control: 293T cells(blue). Primary Antibody: Rabbit Anti-PARK7/CAP1 antibody(SL1306R), Dilution: 0.2μg in 100 μL 1X PBS containing 0.5% BSA; Isotype Control Antibody: Rabbit IgG (orange) ,used under the same conditions. Secondary Antibody: Goat anti-rabbit IgG-PE(white blue), Dilution: 1:200 in 1 X PBS containing 0.5% BSA.
Protocol
Primary antibody (SL1306R, 0.2μg /1x10^6 cells) were incubated for 30 min on the ice, followed by 1 X PBS containing 0.5% BSA + 1 0% goat serum (15 min) to block non-specific protein-protein interactions. Then the Goat Anti-rabbit IgG/PE antibody was added into the blocking buffer mentioned above to react with the primary antibody at 1/200 dilution for 30 min on ice. Acquisition of 20,000 events was performed.
The blue histogram is unstained cells (Hepg2 cells)
concentration 1:50
The Wathet Blue histogram is cells stained with secondary antibody alone.
The Orange histogram is cells stained with rabbit IgG isotype control antibody
plus secondary antibody.
The green histogram is cells stained with Rabbit Anti-PARK7/CAP1 antibody (SL1306R)plus secondary antibody.
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