Peroxiredoxin 3 Recombinant Rabbit Monoclonal Antibody [JA53-21]
Catalog# ET1704-92
Peroxiredoxin 3 Recombinant Rabbit Monoclonal Antibody [JA53-21]
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WB
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IF-Cell
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IHC-P
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FC(Intra)
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IP
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Human
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Rat
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Green monkey
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unconjugated
Overview
Product Name
Peroxiredoxin 3 Recombinant Rabbit Monoclonal Antibody [JA53-21]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within Human Peroxiredoxin 3 aa 207-256 / 256.
Species Reactivity
Human, Rat, Green monkey
Validated Applications
WB, IF-Cell, IHC-P, FC(Intra), IP
Target Molecular Weight
Predicted band size: 28 kDa
Positive Control
HeLa (Human cervical adenocarcinoma cell) cell lysate, LNCaP (Human prostate cancer cell) cell lysate, COS-7 (African green monkey kidney fibroblast) cell lysate, PC-12 (Rat pheochromocytoma cell (undifferentiated)) cell lysate, human liver tissue, human kidney tissue.
Conjugation
unconjugated
Clone Number
JA53-21
RRID
Product Features
Form
Liquid
Concentration
1 mg/mL.(The concentration of this product may be batch-dependent)
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:5,000
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IF-Cell
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1:50-1:200
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IHC-P
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1:1,000
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FC(Intra)
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1:100
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IP
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1-2μg/sample
Target
Function
Thioredoxin-dependent peroxide reductase, mitochondrial is an enzyme that in humans is encoded by the PRDX3 gene. It is a member of the peroxiredoxin family of antioxidant enzymes. This gene encodes a protein with antioxidant function and is localized in the mitochondrion. This gene shows significant nucleotide sequence similarity to the gene coding for the C22 subunit of Salmonella typhimurium alkylhydroperoxide reductase. Expression of this gene product in E. coli deficient in the C22-subunit gene rescued resistance of the bacteria to alkylhydroperoxide. The human and mouse genes are highly conserved, and they map to the regions syntenic between mouse and human chromosomes. Sequence comparisons with recently cloned mammalian homologues suggest that these genes consist of a family that is responsible for regulation of cellular proliferation, differentiation, and antioxidant functions. Two transcript variants encoding two different isoforms have been found for this gene.
Background References
1. Magarin M et al. Embryonic cardiomyocytes can orchestrate various cell protective mechanisms to survive mitochondrial stress. J Mol Cell Cardiol 97:1-14 (2016).
2. Bhatt NM et al. Restoring redox balance enhances contractility in heart trabeculae from type 2 diabetic rats exposed to high glucose. Am J Physiol Heart Circ Physiol 308:H291-302 (2015).
Sequence Similarity
Belongs to the peroxiredoxin family. AhpC/Prx1 subfamily.
Post-translational Modification
Phosphorylated by LRRK2; phosphorylation reduces perodixase activity.; The enzyme can be inactivated by further oxidation of the cysteine sulfenic acid (C(P)-SOH) to sulphinic acid (C(P)-SO2H) and sulphonic acid (C(P)-SO3H) instead of its condensation to a disulfide bond.
Subcellular Location
Mitochondrion.
Synonyms
Antioxidant protein 1 antibody
AOP 1 antibody
AOP-1 antibody
AOP1 antibody
HBC189 antibody
MER5 antibody
MGC104387 antibody
MGC24293 antibody
mitochondrial antibody
peroxiredoxin 3 antibody
ExpandImages
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Western blot analysis of Peroxiredoxin 3 on different lysates with Rabbit anti-Peroxiredoxin 3 antibody (ET1704-92) at 1/5,000 dilution.
Lane 1: HeLa (Human cervical adenocarcinoma cell) cell lysate
Lane 2: LNCaP (Human prostate cancer cell) cell lysate
Lane 3: COS-7 (African green monkey kidney fibroblast) cell lysate
Lane 4: PC-12 (Rat pheochromocytoma cell (undifferentiated)) cell lysate
Lysates/proteins at 20 µg/Lane.
Exposure time: 2 seconds; ECL: K1801
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary antibody: ET1704-92, 1/5,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃
Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature
Predicted band size: 28 kDa
Observed band size: 25 kDa -
☑ Knockdown (KD)
Western blot analysis of Peroxiredoxin 3 on different lysates with Rabbit anti-Peroxiredoxin 3 antibody (ET1704-92) at 1/1,000 dilution.
Lane 1: HAP1-parental cell lysate
Lane 2: HAP1-Peroxiredoxin 3 KD cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 28 kDa
Observed band size: 25 kDa
Exposure time: 6 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1704-92) at 1/1,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Application: Immunocytochemistry (IF-cell)
Species: Human
Sample: HeLa (Human cervix adenocarcinoma epithelial cell)
Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
Antibody dilution buffer: 1% BSA in PBST.
Primary antibody: ET1704-92, 1/100, overnight at 4℃.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.
Counterstain: Beta tubulin (HA601187, Red), 1/100, overnight at 4℃. The nuclear counterstain was DAPI (Blue). -
Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-Peroxiredoxin 3 antibody (ET1704-92) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1704-92) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-Peroxiredoxin 3 antibody (ET1704-92) at 1/1,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1704-92) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Application: Flow Cytometry (Intra)
Species: Human
Sample: HeLa (Human cervix adenocarcinoma epithelial cell)
Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Tween-20, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature.
Antibody dilution buffer: 1x PBS.
Primary antibody: ET1704-92 (1/100, Red) compared with Rabbit IgG Isotype Control (HA722127, Green), 15 minutes at room temperature.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 15 minutes at room temperature. -
Application: Flow Cytometry (Intra)
Species: Rat
Sample: PC-12 (Rat pheochromocytoma cell (undifferentiated))
Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
Permeabilization: 0.1% Tween-20, 15 minutes at room temperature.
Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature.
Antibody dilution buffer: 1x PBS.
Primary antibody: ET1704-92 (1/100, Red) compared with Rabbit IgG Isotype Control (HA722127, Green), 15 minutes at room temperature.
Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 15 minutes at room temperature. -
Immunoprecipitation (IP)
Peroxiredoxin 3 was immunoprecipitated in 0.2 mg HeLa (Human cervix adenocarcinoma epithelial cell) cell lysate with ET1704-92 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET1704-92 at 1/5,000 dilution. Anti-Rabbit IgG for IP, AlpSdAbs® VHH(HRP) (025-100-005) at 1/5,000 dilution was used for 1 hour at room temperature.
Lane 1: HeLa cell lysate (input)
Lane 2: ET1704-92 IP in HeLa cell lysate
Lane 3: Rabbit IgG instead of ET1704-92 in HeLa cell lysate
Exposure time: 1 second
Blocking: 5% NFDM/TBST, 1 hour at room temperature
Primary dilution: ET1704-92, 1/5,000 in primary antibody dilution buffer (K1803), 2 hours at room temperature
Predicted band size: 28 kDa
Observed band size: 25 kDa
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"