AGE ELISA Kit

Produktnummer ABIN6953551

Produktdetails AGE ELISA Kit

Target: AGE (Advanced Glycation End Products (AGE))

Reaktivität: Diverse Spezies

Nachweismethode: Colorimetric

Methodentyp: Competition ELISA

Detektionsbereich: 98.8 ng/mL - 8000 ng/mL

Untere Nachweisgrenze: 98.8 ng/mL

Applikation: ELISA

Verwendungszweck: The kit is a competitive inhibition enzyme immunoassay technique for the in vitro quantitative measurement of AGE in serum, plasma.

Proben: Plasma, Serum

Analytische Methode: Quantitative

Spezifität: This assay has high sensitivity and excellent specificity for detection of Advanced Glycation End Product (AGE)

Sensitivität: 38.2 ng/mL

Bestandteile:

• 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

Anwendungsinformationen

Kommentare:

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.

Probenmenge: 50 μL

Testdauer: 2 h

Plattentyp: Pre-coated

Protokoll:

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

Aufbereitung der Reagenzien:

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 0.7 mL of Standard Diluent, kept for 10 minutes at room temperature, shake gently(not to foam). The concentration of the standard in the stock solution is 8,000 ng/mL. Please prepare 5 tubes containing 0.6 mL Standard Diluent and produce a triple dilution series according to the picture shown below. Mix each tube thoroughly before the next transfer. Set up 5 points of diluted standard such as 8,000 ng/mL, 2666.7 ng/mL, 888.9 ng/mL, 296.3 ng/mL, 98.8 ng/mL, and the last EP tubes 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, kept 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. The standard for this kit is liquid. Due to its small volume, maybe invisible to the eye.
2. Making serial dilution in the wells directly is not permitted.
3. Prepare standard within 15 minutes before assay. Please do not dissolve the reagents at 37 °C directly.
4. Detection Reagent A and B are sticky solutions, therefore, slowly pipette them to reduce the volume errors.
5. 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.
6. The reconstituted Standards, Detection Reagent A and Detection Reagent B can be used only once.
7. If crystals have formed in the Wash Solution concentrate (30x), warm to room temperature and mix gently until the crystals are completely dissolved.
8. Contaminated water or container for reagent preparation will influence the detection result.

Aufbereitung der Proben:

• 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).

Testpräzision: 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%

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

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

Lagerung: 4 °C/-20 °C

Informationen zur Lagerung:

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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Haltbarkeit: 6 months

Referenzen für AGE ELISA Kit (ABIN6953551)

Tang, Yu, Wu, Chen, Zeng, Wang, Yang, Mei, Cao, Qin: "Lychee seed extract protects against neuronal injury and improves cognitive function in rats with type II diabetes mellitus with cognitive impairment." in: International journal of molecular medicine, Vol. 41, Issue 1, pp. 251-263, (2018) (PubMed).

Yang, Meng, Chen, Li, Cen, Wen, Zhang, Li: "Inhibition of Methylglyoxal-Induced AGEs/RAGE Expression Contributes to Dermal Protection by N-Acetyl-L-Cysteine." in: Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology, Vol. 41, Issue 2, pp. 742-754, (2017) (PubMed).

Galiniak, Bartosz, Sadowska-Bartosz: "Glutathione is the main endogenous inhibitor of protein glycation." in: General physiology and biophysics, Vol. 36, Issue 2, pp. 175-186, (2017) (PubMed).

Qiu, Tang, Wei: "Berberine exerts renoprotective effects by regulating the AGEs-RAGE signaling pathway in mesangial cells during diabetic nephropathy." in: Molecular and cellular endocrinology, Vol. 443, pp. 89-105, (2017) (PubMed).

Sun, Huang, Xu, Sun, Lv, Wang: "Advanced glycation end products promote VEGF expression and thus choroidal neovascularization via Cyr61-PI3K/AKT signaling pathway." in: Scientific reports, Vol. 7, Issue 1, pp. 14925, (2017) (PubMed).

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Malmstedt, Frebelius, Lengquist, Jörneskog, Wang, Swedenborg: "The Receptor for Advanced Glycation End Products (Rage) and Its Ligands in Plasma and Infrainguinal Bypass Vein." in: European journal of vascular and endovascular surgery : the official journal of the European Society for Vascular Surgery, Vol. 51, Issue 4, pp. 579-86, (2016) (PubMed).

Dong, Zhang, Liu, Zhu, Zhao, Li, Sun, Ding, Hu, Yang, Zhang, Li: "Beneficial effects of urine-derived stem cells on fibrosis and apoptosis of myocardial, glomerular and bladder cells." in: Molecular and cellular endocrinology, Vol. 427, pp. 21-32, (2016) (PubMed).

Kosutova, Mikolka, Kolomaznik, Balentova, Calkovska, Mokra: "Effects of S-Nitroso-N-Acetyl-Penicillamine (SNAP) on Inflammation, Lung Tissue Apoptosis and iNOS Activity in a Rabbit Model of Acute Lung Injury." in: Advances in experimental medicine and biology, Vol. 935, pp. 13-23, (2016) (PubMed).

Sadowska-Bartosz, Galiniak, Skolimowski, Stefaniuk, Bartosz: "Nitroxides prevent protein glycoxidation in vitro." in: Free radical research, Vol. 49, Issue 2, pp. 113-21, (2015) (PubMed).

Xu, Cai, Sun, Liao, Liu, Xiao, Zhu, Liu, Zhang: "Serum advanced glycation end products are associated with insulin resistance in male nondiabetic patients with obstructive sleep apnea." in: Sleep & breathing = Schlaf & Atmung, (2015) (PubMed).

Zhiping, Imam, Ismail, Ismail, Yida, Ideris, Sarega, Mahmud: "Effects of edible bird's nest on hippocampal and cortical neurodegeneration in ovariectomized rats." in: Food & function, Vol. 6, Issue 5, pp. 1701-11, (2015) (PubMed).

Cai, Sun, Liu, Xu, Xiao, Duan, Liu, Zhang: "Relationship between serum levels of endogenous secretory RAGE and blood pressure in male nondiabetic patients with obstructive sleep apnea." in: Journal of human hypertension, Vol. 29, Issue 12, pp. 713-8, (2015) (PubMed).

Sadowska-Bartosz, Stefaniuk, Galiniak, Bartosz: "Glycation of bovine serum albumin by ascorbate in vitro: Possible contribution of the ascorbyl radical?" in: Redox biology, Vol. 6, pp. 93-9, (2015) (PubMed).

Wu, Li, Li, Sui, Yang, Dong, Xie, Sun: "Inhibition of Advanced Glycation Endproduct Formation by Lotus Seedpod Oligomeric Procyanidins through RAGE-MAPK Signaling and NF-κB Activation in High-Fat-Diet Rats." in: Journal of agricultural and food chemistry, Vol. 63, Issue 31, pp. 6989-98, (2015) (PubMed).

Sadowska-Bartosz, Bartosz: "Ascorbic acid and protein glycation in vitro." in: Chemico-biological interactions, Vol. 240, pp. 154-62, (2015) (PubMed).

Li, Zhang, Chen, Zhang, Ma, Xia: "Establishment of a rabbit model to study the influence of advanced glycation end products accumulation on osteoarthritis and the protective effect of pioglitazone." in: Osteoarthritis and cartilage / OARS, Osteoarthritis Research Society, (2015) (PubMed).

Nikooyeh, Neyestani, Tayebinejad, Alavi-Majd, Shariatzadeh, Kalayi, Zahedirad, Heravifard, Salekzamani: "Daily intake of vitamin D- or calcium-vitamin D-fortified Persian yogurt drink (doogh) attenuates diabetes-induced oxidative stress: evidence for antioxidative properties of vitamin D." in: Journal of human nutrition and dietetics : the official journal of the British Dietetic Association, Vol. 27 Suppl 2, pp. 276-83, (2014) (PubMed).

Mai, Wang, Wen, Lin, Niu: "Lipoprotein-associated phospholipase A2 and AGEs are associated with cardiovascular risk factors in women with history of gestational diabetes mellitus." in: Gynecological endocrinology : the official journal of the International Society of Gynecological Endocrinology, Vol. 30, Issue 3, pp. 241-4, (2014) (PubMed).

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Details zu AGE

Target: AGE (Advanced Glycation End Products (AGE))

Andere Bezeichnung: Advanced Glycation End Product (AGE) (AGE Produkte)

Synonyme: RBP ELISA Kit, RNBP ELISA Kit, renin binding protein ELISA Kit, RENBP ELISA Kit

Substanzklasse: Chemical