PON1 ELISA Kit

Catalog No. ABIN6958390

Product Details PON1 ELISA Kit

Target: PON1 (Paraoxonase 1 (PON1))

Reactivity: Human

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 3.12 ng/mL - 200 ng/mL

Minimum Detection Limit: 3.12 ng/mL

Application: ELISA

Purpose: The kit is a sandwich enzyme immunoassay for in vitro quantitative measurement of PON1 in human serum, plasma, tissue homogenates, cell lysates, cell culture supernates.

Sample Type: Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate

Analytical Method: Quantitative

Specificity: This assay has high sensitivity and excellent specificity for detection of Paraoxonase 1 (PON1)

Sensitivity: 1.27 ng/mL

Components:

• Pre-coated, ready to use 96-well strip plate, flat buttom
• Plate sealer for 96 wells
• Reference Standard
• Standard Diluent
• Detection Reagent A
• Detection Reagent B
• Assay Diluent A
• Assay Diluent B
• Reagent Diluent (if Detection Reagent is lyophilized)
• TMB Substrate
• Stop Solution
• Wash Buffer (30 x concentrate)
• Instruction manual

Application Details

Comment:

Information on standard material:
The standard might be recombinant protein or natural protein, that will depend on the specific kit. Moreover, the expression system is E.coli or yeast or mammal cell. There is 0.05% proclin 300 in the standard as preservative.

Information on reagents:
The stop solution used in the kit is sulfuric acid with concentration of 1 mol/L. And the wash solution is TBS. The standard diluent contains 0.02 % sodium azide, assay diluent A and assay diluent B contain 0.01% sodium azide. Some kits can contain is BSA in them.

Information on antibodies:
The provided antibodies and their host vary in different kits.

Sample Volume: 100 μL

Assay Time: 3 h

Plate: Pre-coated

Protocol:

1. Prepare all reagents, samples and standards,
2. Add 100μL standard or sample to each well. Incubate 1 hours at 37 °C,
3. Aspirate and add 100μL prepared Detection Reagent A. Incubate 1 hour at 37 °C,
4. Aspirate and wash 3 times,
5. Add 100μL prepared Detection Reagent B. Incubate 30 minutes at 37 °C,
6. Aspirate and wash 5 times,
7. Add 90μL Substrate Solution. Incubate 10-20 minutes at 37 °C,
8. Add 50μL Stop Solution. Read at 450nm immediately.

Reagent Preparation:

1. Bring all kit components and samples to room temperature (18-25 °C) before use. If the kit will not be used up in one time, please only take out strips and reagents for present experiment, and leave the remaining strips and reagents in required condition.
2. Standard - Reconstitute the Standard with 1.0 mL of Standard Diluent, keep for 10 minutes at room temperature, shake gently (not to foam). The concentration of the standard in the stock solution is 2,000 ng/mL. Firstly dilute the stock solution to 200 ng/mL and the diluted standard serves as the highest standard (200 ng/mL). Then prepare 7 tubes containing 0.5 mL Standard Diluent and use the diluted standard to produce a double dilution series. Mix each tube thoroughly before the next transfer. Set up 7 points of diluted standard such as 200 ng/mL, 100 ng/mL, 50 ng/mL, 25 ng/mL, 12.5 ng/mL, 6.25 ng/mL, 3.12 ng/mL, and the last microcentrifuge tube with Standard Diluent is the blank as 0 ng/mL.
3. Detection Reagent A and Detection Reagent B - If lyophilized reconstitute the Detection Reagent A with 150μL of Reagent Diluent, keep for 10 minutes at room temperature, shake gently (not to foam). Briefly spin or centrifuge the stock Detection A and Detection B before use. Dilute them to the working concentration 100-fold with Assay Diluent A and B, respectively.
4. Wash Solution - Dilute 20 mL of Wash Solution concentrate (30x) with 580 mL of deionized or distilled water to prepare 600 mL of Wash Solution (1x).
5. TMB substrate - Aspirate the needed dosage of the solution with sterilized tips and do not dump the residual solution into the vial again.

Note:

1. Making serial dilution in the wells directly is not permitted.
2. Prepare standards within 15 minutes before assay. Please do not dissolve the reagents at 37 °C directly.
3. Please carefully reconstitute Standards or working Detection Reagent A and B according to the instruction, and avoid foaming and mix gently until the crystals are completely dissolved. To minimize imprecision caused by pipetting, use small volumes and ensure that pipettors are calibrated. It is recommended to suck more than 10μL for one pipetting.
4. The reconstituted Standards, Detection Reagent A and Detection Reagent B can be used only once.
5. If crystals have formed in the Wash Solution concentrate (30x), warm to room temperature and mix gently until the crystals are completely dissolved.
6. Contaminated water or container for reagent preparation will influence the detection result.

Sample Preparation:

• It is recommended to use fresh samples without long storage, otherwise protein degradation and denaturationmay occur in these samples, leading to false results. Samples should therefore be stored for a short periodat 2 - 8 °C or aliquoted at -20 °C (≤1 month) or -80 °C (≤ 3 months). Repeated freeze-thawcycles should be avoided. Prior to assay, the frozen samples should be slowly thawed and centrifuged toremove precipitates.
• If the sample type is not specified in the instructions, a preliminary test is necessary to determinecompatibility with the kit.
• If a lysis buffer is used to prepare tissue homogenates or cell culture supernatant, there is a possibilityof causing a deviation due to the introduced chemical substance.The recommended dilution factor is for reference only.
• Please estimate the concentration of the samples before performing the test. If the values are not in therange of the standard curve, the optimal sample dilution for the particular experiment has to be determined.Samples should then be diluted with PBS (pH =7.0-7.2).

Assay Precision: Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level of target were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level of target were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV < 10%
Inter-Assay: CV < 12%

Restrictions: For Research Use only

Handling

Precaution of Use: The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.

Storage: 4 °C/-20 °C

Storage Comment:

1. For unopened kit: All reagents should be stored according to the labels on the vials. The Standard, Detection Reagent A, Detection Reagent B, and 96-well Strip Plate should be stored at -20 °C upon receipt, while the other reagents should be stored at 4 °C.
2. For opened kits: the remaining reagents must be stored according to the above storage conditions. In addition, please return the unused wells to the foil pouch containing the desiccant and seal the foil pouch with the zipper.

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Expiry Date: 6 months

References for PON1 ELISA Kit (ABIN6958390)

Pop, Niculae, Pop, Răchișan: "Individuals with autism have higher 8-Iso-PGF2α levels than controls, but no correlation with quantitative assay of Paraoxonase 1 serum levels." in: Metabolic brain disease, Vol. 32, Issue 6, pp. 1943-1950, (2018) (PubMed).

Crow, Meek, Wills, Chambers: "A case-control study: The association of serum paraoxonase 1 activity and concentration with the development of type 2 diabetes mellitus." in: Diabetes/metabolism research and reviews, Vol. 34, Issue 3, (2018) (PubMed).

Singh, Singh, Chandra, Tyagi: "Paraoxonase-1 is a better indicator than HDL of Atherosclerosis - A pilot study in North Indian population." in: Diabetes & metabolic syndrome, Vol. 12, Issue 3, pp. 275-278, (2018) (PubMed).

Ogłodek: "The role of PON-1, GR, IL-18, and OxLDL in depression with and without posttraumatic stress disorder." in: Pharmacological reports : PR, Vol. 69, Issue 5, pp. 837-845, (2018) (PubMed).

Samouilidou, Bountou, Papandroulaki, Papamanolis, Papakostas, Grapsa: "Serum Endocan Levels are Associated With Paraoxonase 1 Concentration in Patients With Chronic Kidney Disease." in: Therapeutic apheresis and dialysis : official peer-reviewed journal of the International Society for Apheresis, the Japanese Society for Apheresis, the Japanese Society for Dialysis Therapy, Vol. 22, Issue 4, pp. 325-331, (2018) (PubMed).

Samouilidou, Kostopoulos, Liaouri, Kioussi, Vassiliou, Bountou, Grapsa: "Association of lipid profile with serum PON1 concentration in patients with chronic kidney disease." in: Renal failure, pp. 1-6, (2016) (PubMed).

Turgut Cosan, Colak, Saydam, Yazıcı, Degirmenci, Birdane, Colak, Gunes: "Association of paraoxonase 1 (PON1) gene polymorphisms and concentration with essential hypertension." in: Clinical and experimental hypertension (New York, N.Y. : 1993), Vol. 38, Issue 7, pp. 602-607, (2016) (PubMed).

Johansson, Bro-Jeppesen, Kjaergaard, Wanscher, Hassager, Ostrowski et al.: "Sympathoadrenal activation and endothelial damage are inter correlated and predict increased mortality in patients resuscitated after out-of-hospital cardiac arrest. a post Hoc sub-study of patients ..." in: PLoS ONE, Vol. 10, Issue 3, pp. e0120914, (2015) (PubMed).

Taylor, Plaisance, Mahurin, Mestek, Moncada-Jimenez, Grandjean: "Paraoxonase responses to exercise and niacin therapy in men with metabolic syndrome." in: Redox report : communications in free radical research, Vol. 20, Issue 1, pp. 42-8, (2014) (PubMed).

Loued, Berrougui, Componova, Ikhlef, Helal, Khalil: "Extra-virgin olive oil consumption reduces the age-related decrease in HDL and paraoxonase 1 anti-inflammatory activities." in: The British journal of nutrition, Vol. 110, Issue 7, pp. 1272-84, (2013) (PubMed).

Patra, Singh, Singh: "Paraoxonase 1: a better atherosclerotic risk predictor than HDL in type 2 diabetes mellitus." in: Diabetes & metabolic syndrome, Vol. 7, Issue 2, pp. 108-11, (2013) (PubMed).

Oberbach, von Bergen, Blüher, Lehmann, Till: "Combined serum proteomic and metabonomic profiling after laparoscopic sleeve gastrectomy in children and adolescents." in: Journal of laparoendoscopic & advanced surgical techniques. Part A, Vol. 22, Issue 2, pp. 184-8, (2012) (PubMed).

Burillo, Mateo-Gallego, Cenarro, Fiddyment, Bea, Jorge, Vázquez, Civeira: "Beneficial effects of omega-3 fatty acids in the proteome of high-density lipoprotein proteome." in: Lipids in health and disease, Vol. 11, Issue 1, pp. 116, (2012) (PubMed).

Liang, Lee, Lee, Lee, Lin, Hsu, Wan, Yu, Tsai, Fu, Ting, Sheu: "Persistent elevation of paraoxonase-1 specific enzyme activity after weight reduction in obese non-diabetic men with metabolic syndrome." in: Clinica chimica acta; international journal of clinical chemistry, Vol. 412, Issue 19-20, pp. 1835-41, (2011) (PubMed).

Yetukuri, Huopaniemi, Koivuniemi, Maranghi, Hiukka, Nygren, Kaski, Taskinen, Vattulainen, Jauhiainen, Oreši?: "High density lipoprotein structural changes and drug response in lipidomic profiles following the long-term fenofibrate therapy in the FIELD substudy." in: PLoS ONE, Vol. 6, Issue 8, pp. e23589, (2011) (PubMed).

Maranghi, Hiukka, Badeau, Sundvall, Jauhiainen, Taskinen: "Macrophage cholesterol efflux to plasma and HDL in subjects with low and high homocysteine levels: a FIELD substudy." in: Atherosclerosis, Vol. 219, Issue 1, pp. 259-65, (2011) (PubMed).

Er?ahin, Toklu, Erzik, Cetinel, Akakin, Velio?lu-O?ünç, Tetik, Ozdemir, Sener, Ye?en: "The anti-inflammatory and neuroprotective effects of ghrelin in subarachnoid hemorrhage-induced oxidative brain damage in rats." in: Journal of neurotrauma, Vol. 27, Issue 6, pp. 1143-55, (2010) (PubMed).

Kim, Kobashi: "Immobilized arylsulfotransferase." in: Journal of biochemistry, Vol. 102, Issue 3, pp. 487-91, (1988) (PubMed).

Target Details for PON1

Target: PON1 (Paraoxonase 1 (PON1))

Abstract: PON1 Products

Synonyms: esa ELISA Kit, pon ELISA Kit, pon2 ELISA Kit, MGC53915 ELISA Kit, MGC89222 ELISA Kit, PON1 ELISA Kit, ESA ELISA Kit, MVCD5 ELISA Kit, PON ELISA Kit, Pon ELISA Kit, paraoxonase 2 ELISA Kit, paraoxonase 1 ELISA Kit, pon2 ELISA Kit, PON1 ELISA Kit, Pon1 ELISA Kit