ET1704-92

Peroxiredoxin 3 Recombinant Rabbit Monoclonal Antibody [JA53-21]

cat.: ET1704-92

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Rat, Green monkey
Applications:	WB, IF-Cell, IHC-P, FC(Intra), IP
Clonality:	Monoclonal
Clone number:	JA53-21

Form:	Liquid
Storage condition:	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.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 28 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human Peroxiredoxin 3 aa 207-256 / 256.
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.
Subcellular location:	Mitochondrion.
Recommended Dilutions: WB IF-Cell IHC-P FC(Intra) IP	1:5,000 1:50-1:200 1:1,000 1:100 1-2μg/sample
Uniprot #:	SwissProt: P30048 Human \| Q9Z0V6 Rat
Alternative names:	Antioxidant protein 1 AOP 1 AOP-1 AOP1 HBC189 MER5 MGC104387 MGC24293 mitochondrial peroxiredoxin 3 Peroxiredoxin III Peroxiredoxin-3 PRDX3 PRDX3_HUMAN PRO1748 Protein MER5 homolog PRX III Prx-III PRX3 SP 22 SP-22 SP22 Thioredoxin dependent peroxide reductase mitochondrial Thioredoxin-dependent peroxide reductase

Images

Fig1: 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

Fig2: 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.

	Fig3: 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).
	Fig4: 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.
	Fig5: 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.

Fig6: 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.

Fig7: 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.

Fig8: 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

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.