Anti-Peroxiredoxin 1/PRDX1 Antibody Picoband®

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.

Prdx1 expression was significantly increased in rats after ICH. (A) Prdx1 expression in the perihematomal area was detected by qRT-PCR (df = 4, F = 35.945, ∗ P < 0.05 versus sham, ∗∗ P < 0.01 versus sham, n = 3). (B) Survival statistics of the sham group and WT group (χ2 = 10.457, df = 1, ∗∗ P = 0.01, n = 20). (C) Prdx1 expression in the perihematomal area was detected by western blot analysis ( F = 2.014, t = –3.432, ∗∗ P < 0.01 versus sham, n = 3). (D) Prdx1 expression in perihematomal tissue was detected by immunohistochemistry (scale bars, 100 μm, F = 1.225, t = –7.288, ∗∗ P < 0.01 versus sham, n = 3). (E) Prdx1 expression in perihematomal tissue was determined by immunofluorescence, and the percentage of positive Prdx1/NeuN, Prdx1/Ibα-1 and Prdx1/GFAP in three randomly chosen fields within the perihematomal area was counted (scale bars, 25 μm, F = 10.964, t = –11.790, ## P < 0.01 versus Prdx1/NeuN. F = 12.000, t = –60.228, ∗∗ P < 0.01 versus Prdx1/NeuN, n = 3).
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Prdx1 overexpression alleviated the symptoms of rats after ICH. (A) Survival statistics of the sham group, WT group, Vector group, and Prdx1-OE group (χ2 = 14.310, df = 3, ∗ P < 0.05 versus Vector, # P < 0.05 versus WT, n = 20). (B) Brain water content of the four groups (df = 3, F = 9.324, ∗ P < 0.05 versus Vector, ## P < 0.01 versus WT, n = 6). (C) Hematoma volume of the sham group, the WT group, the Vector group, and the Prdx1-OE group (df = 3, F = 151.467, ∗ P < 0.05 versus Vector, # P < 0.05 versus WT, n = 6). (D) The mNSS was determined for four groups 3 days after ICH (df = 3, F = 75.196, ∗∗ P < 0.01 versus Vector, ## P < 0.01 versus WT, n = 5).
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Prdx1 overexpression inhibited inflammation and apoptosis after ICH. (A) Representative Nissl staining in sham, WT, Prdx1-OE and Vector rats. The number of Nissl-positive cells was assessed (scale bars, 50 μm, df = 3, F = 174.206, ∗∗ P < 0.01 versus Vector; ## P < 0.01 versus WT, n = 3). (B) Representative FJB staining in sham, WT, Prdx1-OE, and Vector rats. The number of FJB-positive cells was assessed (scale bars, 50 μm, df = 3, F = 435.050, ∗∗ P < 0.01 versus Vector; ## P < 0.01 versus WT, n = 3). (C) Western blotting was used to detect Bcl2 and Bax expression in the four groups (df = 3, F = 32.759, ∗ P < 0.05 versus Vector; # P < 0.05 versus WT, n = 3). (D) Comparison of the expression of inflammatory factors in the four groups using qRT-PCR: IL-6 (df = 3, F = 27.046, ∗ P < 0.05 versus Vector; ## P < 0.01 versus WT, n = 3), IL-10 (df = 3, F = 79.041, ∗∗ P < 0.01 versus Vector, ## P < 0.01 versus WT, n = 3), TNF-α (df = 3, F = 10.274, ∗∗ P < 0.01 versus Vector; # P < 0.05 versus WT, n = 3).
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The genome-wide landscape of Prdx1 binding sites on RNA. (A) Venn diagram of Prdx1 RIP-seq genes from two biological replicates. (B) Heat map showing the correlation coefficient of two biological replicates. (C) The Prdx1 RIP-seq peaks are predominantly enriched in the CDS region, the 3′ UTR and the 5′ UTR. All RIP-seq peaks were classified according to their distribution on the RNA elements and compared to the genomic background. (D) De novo motif analysis identified GA repeat and GA-enriched sequences as Prdx1 binding motifs. (E) Prdx1 RIP-seq peak distribution proportion. (F) Prdx1 RIP-seq peaks are shown as track signals. The peak area is indicated by the black frame. (G) RIP-qPCR analysis of Prdx1 binding RNAs, IgG RIP was negative RIP control (BCL6: F = 3.745, t = –6.708, ∗∗ P < 0.01 versus IgG, n = 3; TLR6: F = 1.668, t = –7.065, ∗∗ P < 0.01 versus IgG, n = 3; FOS: F = 5.224, t = –10.432, ∗∗ P < 0.01 versus IgG, n = 3; PTEN: F = 0.000, t = –6.998, ∗∗ P < 0.01 versus IgG, n = 3).
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Prdx1 affects inflammation- and apoptosis-related mRNA stability. (A) Prdx1 expression in control HeLa cells and in the Prdx1-OE group was detected by qRT-PCR ( F = 8.494, t = 40.635, ∗∗ P < 0.01 versus Control). (B) Scatter plots and correlation coefficients of two biological replicates of RNA-seq. (C) Volcano map: red dot indicates a gene that is upregulated after Prdx1 overexpression, black dot indicates a gene with no significant change, and blue dot indicates a downregulated gene. (D) Gene ontology (GO) analysis of Prdx1-dependent DEGs. Significantly enriched GO terms of genes. The x -axis indicates the enrichment P -value on a −log10 scale; the y -axis indicates terms. (E) Heat map showing that apoptosis- and inflammation-related genes were significantly decreased after Prdx1 was upregulated in HeLa cells. (F) qRT-PCR was performed in four groups 3 days after ICH (BMP2: F = 8.949, df = 3, ∗ P < 0.05 versus Vector, # P < 0.05 versus WT, n = 3; CCL2: F = 33.103, df = 3, ∗∗ P < 0.01 versus Vector, ## P < 0.01 versus WT, n = 3; TLR3: F = 39.330, df = 3, ∗∗ P < 0.01 versus Vector, ## P < 0.01 versus WT, n = 3).
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Prdx1 plays similar roles in vitro and in vivo . (A) Prdx1 RIP-seq peaks are shown as track signals of ANGPTL4, GADD45A and THBS1. The peak area is indicated by the black frame. (B) RIP-qPCR analysis of Prdx1 binding RNAs in astrocytes with IgG RIP as a negative RIP control (ANGPTL4: F = 4.297, t = –13.551, ∗∗ P < 0.01 versus IgG, n = 3; GADD45A: F = 12.277, t = –8.909, ∗∗ P < 0.01 versus IgG, n = 3; THBS: F = 4.072, t = –20.619, ∗∗ P < 0.01 versus IgG, n = 3). (C) qRT-PCR was performed in four groups of ANGPTL4, GADD45A, and THBS1 mRNA (ANGPTL4: F = 87.049, df = 3, ∗ P < 0.05 versus Vector, # P < 0.05 versus WT, n = 3; GADD45A: F = 73.252, df = 3, ∗∗ P < 0.01 versus Vector, ## P < 0.01 versus WT, n = 3; THBS: F = 20.553, df = 3, ∗∗ P < 0.01 versus Vector, ## P < 0.01 versus WT, n = 3).
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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.

Flow Cytometry analysis of HEPG2 cells using anti-Peroxiredoxin 1 antibody (PB9348).
Overlay histogram showing HEPG2 cells stained with PB9348 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. 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 without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

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.

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.

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.

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.

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.

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.