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Rat PRDX5 / Peroxiredoxin-5 ELISA Kit

SKU:
RTFI00232
Product Type:
ELISA Kit
Size:
96 Assays
Uniprot:
Q9R063
Sensitivity:
0.469ng/ml
Range:
0.781-50ng/ml
ELISA Type:
Sandwich
Synonyms:
Prdx5, ACR1, AOEB166, AOPP, PMP20, PRXV, Peroxisomal antioxidant enzyme, Alu corepressor 1, Alu co-repressor 1, Antioxidant enzyme B166, Peroxiredoxin V, B166, Liver tissue 2D-page spot 71B, peroxiredoxin 5, peroxiredoxin-5, mitochondrial, PLPThiored
Reactivity:
Rat
$779
Frequently bought together:

Description

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Rat PRDX5/Peroxiredoxin-5 ELISA Kit

The Rat PRDX5 (Peroxiredoxin-5) ELISA Kit is a powerful tool for detecting levels of peroxiredoxin-5 in rat serum, plasma, and cell culture supernatants with precision and accuracy. This kit offers exceptional sensitivity and specificity, guaranteeing dependable and consistent results for a variety of research applications.Peroxiredoxin-5 is a vital antioxidant enzyme that plays a critical role in protecting cells from oxidative stress and maintaining cellular homeostasis.

Its involvement in various biological processes makes it a key molecule to study in diseases related to oxidative damage, such as cancer, neurodegenerative disorders, and cardiovascular diseases. The Rat PRDX5 ELISA Kit provides researchers with a reliable means to measure peroxiredoxin-5 levels, aiding in the understanding of disease mechanisms and the development of potential therapeutic interventions.

Product Name:Rat Prdx5 (Peroxiredoxin-5, mitochondrial) ELISA Kit
Product Code:RTFI00232
Size:96 Assays
Target:Rat Prdx5
Alias:Prdx5, ACR1, AOEB166, AOPP, PMP20, PRXV, Peroxisomal antioxidant enzyme, Alu corepressor 1, Alu co-repressor 1, Antioxidant enzyme B166, Peroxiredoxin V, B166, Liver tissue 2D-page spot 71B, peroxiredoxin 5, peroxiredoxin-5, mitochondrial, PLPThioredoxin peroxidase PMP20, PRDX6, prx-V, SBBI10, Thioredoxin reductase, TPx type VI
Reactivity:Rat
Detection Method:Sandwich ELISA, Double Antibody
Sensitivity:0.469ng/ml
Range:0.781-50ng/ml
Storage:4°C for 6 months
Note:For Research Use Only
Recovery:Matrices listed below were spiked with certain level of Rat Prdx5 and the recovery rates were calculated by comparing the measured value to the expected amount of Rat Prdx5 in samples.
MatrixRecovery range(%)Average(%)
serum(n=5)85-10595
EDTA plasma(n=5)86-10397
UFH plasma(n=5)88-10397
Linearity:The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Rat Prdx5 and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample1:21:41:8
serum(n=5)89-95%90-101%85-101%
EDTA plasma(n=5)82-101%84-97%82-91%
UFH plasma(n=5)88-99%80-97%85-100%
Intra-Assay:CV <8%
Inter-Assay:CV <10%
Uniprot:Q9R063
UniProt Protein Function:PRDX5: Reduces hydrogen peroxide and alkyl hydroperoxides with reducing equivalents provided through the thioredoxin system. Involved in intracellular redox signaling. Belongs to the peroxiredoxin 2 family. 2 isoforms of the human protein are produced by alternative initiation.
UniProt Protein Details:

Protein type:Oxidoreductase; EC 1.11.1.15

Cellular Component: cytoplasm; cytoplasmic vesicle; cytosol; extracellular space; intracellular membrane-bound organelle; mitochondrion; nucleus; perinuclear region of cytoplasm; peroxisomal matrix; peroxisome

Molecular Function:antioxidant activity; caspase inhibitor activity; peroxidase activity; peroxiredoxin activity; protein dimerization activity; receptor binding

Biological Process: apoptosis; hydrogen peroxide catabolic process; negative regulation of apoptosis; negative regulation of caspase activity; negative regulation of oxidoreductase activity; negative regulation of transcription from RNA polymerase III promoter; positive regulation of collagen biosynthetic process; response to oxidative stress

NCBI Summary:member of the AhpC/TSA peroxiredoxin family with mitochondrial and peroxisomal sorting signals; may play an antioxidant protective role in various tissues [RGD, Feb 2006]
UniProt Code:Q9R063
NCBI GenInfo Identifier:16758404
NCBI Gene ID:113898
NCBI Accession:NP_446062.1
UniProt Secondary Accession:Q9R063,Q68G22,
UniProt Related Accession:Q9R063
Molecular Weight:17,035 Da
NCBI Full Name:peroxiredoxin-5, mitochondrial
NCBI Synonym Full Names:peroxiredoxin 5
NCBI Official Symbol:Prdx5  
NCBI Official Synonym Symbols:Aoeb166  
NCBI Protein Information:peroxiredoxin-5, mitochondrial
UniProt Protein Name:Peroxiredoxin-5, mitochondrial
UniProt Synonym Protein Names:Antioxidant enzyme B166; AOEB166; PLP; Peroxiredoxin V; Prx-V; Peroxisomal antioxidant enzyme; Thioredoxin peroxidase PMP20; Thioredoxin reductase
Protein Family:Peroxiredoxin
UniProt Gene Name:Prdx5  
UniProt Entry Name:PRDX5_RAT
StepProcedure
1. Set standard, test sample and control (zero) wells on the pre-coated plate respectively, and then, record their positions. It is recommended to measure each standard and sample in duplicate. Wash plate 2 times before adding standard, sample and control (zero) wells!
2.Aliquot 0.1ml standard solutions into the standard wells.
3. Add 0.1 ml of Sample / Standard dilution buffer into the control (zero) well.
4. Add 0.1 ml of properly diluted sample ( Human serum, plasma, tissue homogenates and other biological fluids.) into test sample wells.
5.Seal the plate with a cover and incubate at 37°C for 90 min.
6. Remove the cover and discard the plate content, clap the plate on the absorbent filter papers or other absorbent material. Do NOT let the wells completely dry at any time. Wash plate X2.
7. Add 0.1 ml of Biotin- detection antibody working solution into the above wells (standard, test sample & zero wells). Add the solution at the bottom of each well without touching the side wall.
8.Seal the plate with a cover and incubate at 37°C for 60 min.
9. Remove the cover, and wash plate 3 times with Wash buffer. Let wash buffer rest in wells for 1 min between each wash.
10. Add 0.1 ml of SABC working solution into each well, cover the plate and incubate at 37°C for 30 min.
11. Remove the cover and wash plate 5 times with Wash buffer, and each time let the wash buffer stay in the wells for 1-2 min.
12. Add 90 µL of TMB substrate into each well, cover the plate and incubate at 37°C in dark within 10-20 min. (Note: This incubation time is for reference use only, the optimal time should be determined by end user.) And the shades of blue can be seen in the first 3-4 wells (with most concentrated standard solutions), the other wells show no obvious color.
13. Add 50 µL of Stop solution into each well and mix thoroughly. The color changes into yellow immediately.
14. Read the O.D. absorbance at 450 nm in a microplate reader immediately after adding the stop solution.

When carrying out an ELISA assay it is important to prepare your samples in order to achieve the best possible results. Below we have a list of procedures for the preparation of samples for different sample types.

Sample Type Protocol
Serum:

If using serum separator tubes, allow samples to clot for 30 minutes at room temperature. Centrifuge for 10 minutes at 1,000x g. Collect the serum fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.

If serum separator tubes are not being used, allow samples to clotovernight at 2-8°C. Centrifuge for 10 minutes at 1,000x g. Removeserum and assay promptly or aliquot and store the samples at-80°C. Avoid multiple freeze-thaw cycles.

Plasma: Collect plasma using EDTA or heparin as an anti-coagulant. Centrifuge samples at 4°C for 15 mins at 1000 × g within 30 mins of collection. Collect the plasma fraction and assay promptly or aliquot and store the samples at -80°C. Avoid multiple freeze-thaw cycles.Note: Over haemolysed samples are not suitable for use with this kit.
Urine & Cerebrospinal Fluid: Collect the urine (mid-stream) in a sterile container, centrifuge for 20 mins at 2000-3000 rpm. Remove supernatant and assay immediately. If any precipitation is detected, repeat the centrifugation step. A similar protocol can be used for cerebrospinal fluid.
Cell Culture Supernatant: Collect the cell culture media by pipette, followed by centrifugation at 4°C for 20 mins at 1500 rpm. Collect the clear supernatant and assay immediately.
Cell Lysates: Solubilize cells in lysis buffer and allow to sit on ice for 30 minutes. Centrifuge tubes at 14,000 x g for 5 minutes to remove insoluble material. Aliquot the supernatant into a new tube and discard the remaining whole cell extract. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20°C.
Tissue Homogenates: The preparation of tissue homogenates will vary depending upon tissue type. Rinse tissue with 1X PBS to remove excess blood & homogenizein 20ml of 1X PBS (including protease inhibitors) and store overnight at ≤ -20°C. Two freeze-thaw cycles are required to break the cell membranes. To further disrupt the cell membranes you can sonicate the samples. Centrifuge homogenates for 5 mins at 5000xg. Remove the supernatant and assay immediately or aliquot and store at -20°C or-80°C.
Tissue Lysates: Rinse tissue with PBS, cut into 1-2 mm pieces, and homogenize with a tissue homogenizer in PBS. Add an equal volume of RIPA buffer containing protease inhibitors and lyse tissues at room temperature for 30 minutes with gentle agitation. Centrifuge to remove debris. Quantify total protein concentration using a total protein assay. Assay immediately or aliquot and store at ≤ -20 °C.
Breast Milk: Collect milk samples and centrifuge at 10,000 x g for 60 min at 4°C. Aliquot the supernatant and assay. For long term use, store samples at -80°C. Minimize freeze/thaw cycles.

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