Anti-Peroxiredoxin 1/PRDX1 Antibody Picoband™

Figure 1. Western blot analysis of Peroxiredoxin 1 using anti-Peroxiredoxin 1 antibody (PB9348).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human Raji whole cell lysates,
Lane 2: human Jurkat whole cell lysates,
Lane 3: human MDA-MB-453 whole cell lysates,
Lane 4: human Hela whole cell lysates,
Lane 5: human HepG2 whole cell lysates,
Lane 6: human HEK293 whole cell lysates,
Lane 7: human placenta tissue lysates,
Lane 8: human A549 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Peroxiredoxin 1 antigen affinity purified polyclonal antibody (Catalog # PB9348) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Peroxiredoxin 1 at approximately 24 kDa. The expected band size for Peroxiredoxin 1 is at 22 kDa.

Figure 2. Western blot analysis of Peroxiredoxin 1 using anti-Peroxiredoxin 1 antibody (PB9348).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: rat kidney tissue lysates,
Lane 2: rat testis tissue lysates,
Lane 3: rat liver tissue lysates,
Lane 4: rat PC-12 whole cell lysates,
Lane 5: mouse kidney tissue lysates,
Lane 6: mouse testis tissue lysates,
Lane 7: mouse liver tissue lysates,
Lane 8: mouse RAW264.7 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Peroxiredoxin 1 antigen affinity purified polyclonal antibody (Catalog # PB9348) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Peroxiredoxin 1 at approximately 24 kDa. The expected band size for Peroxiredoxin 1 is at 22 kDa.

Figure 3. IHC analysis of Peroxiredoxin 1 using anti-Peroxiredoxin 1 antibody (PB9348).
Peroxiredoxin 1 was detected in a paraffin-embedded section of Human Mammary Cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-Peroxiredoxin 1 Antibody (PB9348) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

Figure 4. IHC analysis of Peroxiredoxin 1 using anti-Peroxiredoxin 1 antibody (PB9348).
Peroxiredoxin 1 was detected in a paraffin-embedded section of Mouse Brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-Peroxiredoxin 1 Antibody (PB9348) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

Figure 5. IHC analysis of Peroxiredoxin 1 using anti-Peroxiredoxin 1 antibody (PB9348).
Peroxiredoxin 1 was detected in a paraffin-embedded section of Rat Brain tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 μg/ml rabbit anti-Peroxiredoxin 1 Antibody (PB9348) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

Figure 6. IHC analysis of Peroxiredoxin 1 using anti-Peroxiredoxin 1 antibody (PB9348).
Peroxiredoxin 1 was detected in immunocytochemical section of SMMC-7721 Cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 1μg/ml rabbit anti-Peroxiredoxin 1 Antibody (PB9348) overnight at 4°C. Biotinylated goat anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The section was developed using Strepavidin-Biotin-Complex (SABC)(Catalog # SA1022) with DAB as the chromogen.

Figure 7. IF analysis of Peroxiredoxin 1 using anti-Peroxiredoxin 1 antibody (PB9348).
Peroxiredoxin 1 was detected in immunocytochemical section of Hela cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL rabbit anti-Peroxiredoxin 1 Antibody (PB9348) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Figure 8. IF analysis of Peroxiredoxin 1 using anti- Peroxiredoxin 1 antibody (PB9348).
Peroxiredoxin 1 was detected in immunocytochemical section of U20S cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL rabbit anti-Peroxiredoxin 1 Antibody (PB9348) overnight at 4°C. DyLight®594 Conjugated Goat Anti-Rabbit IgG (BA1142) was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

Figure 9. Flow Cytometry analysis of HEPG2 cells using anti-Peroxiredoxin 1 antibody (PB9348).
Overlay histogram showing HEPG2 cells stained with PB9348 (Blue line).The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Peroxiredoxin 1 Antibody (PB9348, 1μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.