NUCB2 ELISA Kit

Produktnummer ABIN6958069

Produktdetails NUCB2 ELISA Kit

Target: NUCB2 (NES1) (Nesfatin-1 (NES1))

Reaktivität: Human

Nachweismethode: Colorimetric

Methodentyp: Competition ELISA

Detektionsbereich: 617.3 pg/mL - 50000 pg/mL

Untere Nachweisgrenze: 617.3 pg/mL

Applikation: ELISA

Verwendungszweck: The kit is a competitive inhibition enzyme immunoassay technique for the in vitro quantitative measurement of NES1 in human serum, plasma.

Proben: Plasma, Serum

Analytische Methode: Quantitative

Spezifität: This assay has high sensitivity and excellent specificity for detection of Nesfatin 1 (NES1)

Sensitivität: 234.2 pg/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 1.0 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 50,000pg/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 50,000pg/mL, 16,666.7pg/mL, 5,555.6pg/mL, 1,851.9pg/mL, 617.3pg/mL, and the last EP tubes with Standard Diluent is the blank as 0pg/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. Making serial dilution in the wells directly is not permitted.
2. Prepare standard within 15 minutes before assay. Please do not dissolve the reagents at 37 °C directly.
3. Detection Reagent A and B are sticky solutions, therefore, slowly pipette them to reduce the volume errors.
4. 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.
5. The reconstituted Standards, Detection Reagent A and Detection Reagent B can be used only once.
6. If crystals have formed in the Wash Solution concentrate (30x), warm to room temperature and mix gently until the crystals are completely dissolved.
7. 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 NUCB2 ELISA Kit (ABIN6958069)

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Ramesh, Gawli, Pasupulleti, Unniappan: "Metabolic and Cardiovascular Actions of Nesfatin-1: Implications in Health and Disease." in: Current pharmaceutical design, Vol. 23, Issue 10, pp. 1453-1464, (2018) (PubMed).

Ademoglu, Gorar, Keskin, Carlioglu, Ucler, Erdamar, Culha, Aral: "Serum nesfatin-1 levels are decreased in pregnant women newly diagnosed with gestational diabetes." in: Archives of endocrinology and metabolism, Vol. 61, Issue 5, pp. 455-459, (2018) (PubMed).

Lu, Wang, Tang, Cheng, Du, Wang, Feng, Li, Cai, Qiu, Sun: "Nesfatin-1 functions as a switch for phenotype transformation and proliferation of VSMCs in hypertensive vascular remodeling." in: Biochimica et biophysica acta. Molecular basis of disease, Vol. 1864, Issue 6 Pt A, pp. 2154-2168, (2018) (PubMed).

Ustabaş Kahraman, Vehapoğlu, Özgen, Terzioğlu, Cesur, Dündaröz: "Correlation of Brain Neuropeptide (Nesfatin-1 and Orexin-A) Concentrations with Anthropometric and Biochemical Parameters in Malnourished Children." in: Journal of clinical research in pediatric endocrinology, Vol. 7, Issue 3, pp. 197-202, (2016) (PubMed).

Cakir, Calikoglu, Yılmaz, Akpinar, Bayraktutan, Topcu: "Serum nesfatin-1 levels: a potential new biomarker in patients with subarachnoid hemorrhage." in: The International journal of neuroscience, pp. 1-7, (2016) (PubMed).

Ahmadizad, Avansar, Ebrahim, Avandi, Ghasemikaram: "The effects of short-term high-intensity interval training vs. moderate-intensity continuous training on plasma levels of nesfatin-1 and inflammatory markers." in: Hormone molecular biology and clinical investigation, Vol. 21, Issue 3, pp. 165-73, (2015) (PubMed).

Ülger, Gül, Uslukaya, O?uz, Bozda?, Yüksel, Böyük: "New hormones to predict the severity of gallstone-induced acute pancreatitis." in: The Turkish journal of gastroenterology : the official journal of Turkish Society of Gastroenterology, Vol. 25, Issue 6, pp. 714-7, (2015) (PubMed).

Alp, Görmüş, Güdücü, Bozkurt: "Nesfatin-1 levels and metabolic markers in polycystic ovary syndrome." in: Gynecological endocrinology : the official journal of the International Society of Gynecological Endocrinology, Vol. 31, Issue 7, pp. 543-7, (2015) (PubMed).

Araz, Yilmazel Ucar, Dorman, Bayraktutan, Yayla, Yilmaz, Acemoglu, Halici, Akgun: "Is There a Relationship between Obstructive Sleep Apnea Syndrome Severity and Nesfatin-1?" in: Respiration; international review of thoracic diseases, Vol. 90, Issue 2, pp. 105-10, (2015) (PubMed).

Sahin, Sahin, Ural, Cure, Senturk, Tekin, Balik, Cure, Yuce, Kirbas: "Nesfatin-1 and Vitamin D levels may be associated with systolic and diastolic blood pressure values and hearth rate in polycystic ovary syndrome." in: Bosnian journal of basic medical sciences / Udruženje basičnih mediciniskih znanosti = Association of Basic Medical Sciences, Vol. 15, Issue 3, pp. 57-63, (2015) (PubMed).

?engül, Dilbaz, Hal?c?, Ferah, Çad?rc?, Y?lmaz: "Decreased serum nesfatin-1 levels in endometriosis." in: European journal of obstetrics, gynecology, and reproductive biology, Vol. 177, pp. 34-7, (2014) (PubMed).

Ademoglu, Gorar, Carl?oglu, Yaz?c?, Dellal, Berberoglu, Akdeniz, Uysal, Karakurt: "Plasma nesfatin-1 levels are increased in patients with polycystic ovary syndrome." in: Journal of endocrinological investigation, Vol. 37, Issue 8, pp. 715-9, (2014) (PubMed).

Türko?lu, Böyük, Tanr?verdi, Gündüz, Dusak, Kaplan, Gümü?: "The potential role of BMI, plasma leptin, nesfatin-1 and ghrelin levels in the early detection of pancreatic necrosis and severe acute pancreatitis: a prospective cohort study." in: International journal of surgery (London, England), Vol. 12, Issue 12, pp. 1310-3, (2014) (PubMed).

Aslan, Celik, Celik, Turkcuoglu, Yilmaz, Karaer, Simsek, Celik, Aydin: "Cord blood nesfatin-1 and apelin-36 levels in gestational diabetes mellitus." in: Endocrine, Vol. 41, Issue 3, pp. 424-9, (2012) (PubMed).

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Umut, Evren, Cansiz, Akkus, Karamustafalioglu: "Serum NUCB2/nesfatin-1 levels in different stages of alcohol dependence: Is there a relationship with craving?" in: Indian journal of psychiatry, Vol. 59, Issue 1, pp. 94-99 (PubMed).

Details zu NUCB2

Target: NUCB2 (NES1) (Nesfatin-1 (NES1))

Andere Bezeichnung: Nesfatin 1 (NES1) (NES1 Produkte)

Synonyme: nucleobindin-2 ELISA Kit, nucleobindin 2 ELISA Kit, NUCB2 ELISA Kit, Nucb2 ELISA Kit