Hepcidin Kit ELISA

N° du produit ABIN7059992

Détail du produit Hepcidin Kit ELISA

Antigène: Hepcidin (HAMP) (Hepcidin Antimicrobial Peptide (HAMP))

Reactivité: Souris

Méthode de détection: Colorimetric

Type de méthode: Sandwich ELISA

Gamme de detection: 62.5 pg/mL - 4000 pg/mL

Seuil minimal de détection: 62.5 pg/mL

Application: ELISA

Fonction: The kit is a sandwich enzyme immunoassay for in vitro quantitative measurement of Hepc in mouse serum, plasma, urine.

Type d'échantillon: Plasma, Serum, Urine

Analytical Method: Quantitative

Specificité: This assay has high sensitivity and excellent specificity for detection of Hepcidin (Hepc)

Sensibilité: 25.2 pg/mL

Ingrédients:

• 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

Information d'application

Commentaires:

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.

Volume d'échantillon: 100 μL

Durée du test: 3 h

Plaque: Pre-coated

Protocole:

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.

Préparation des réactifs:

1. Bring all kit components and samples to room temperature (18-25 °C) before use.
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 40,000pg/mL. Firstly dilute the stock solution to 4,000pg/mL and the diluted standard serves as the highest standard (4,000pg/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 4,000pg/mL, 2,000pg/mL, 1,000pg/mL, 500pg/mL, 250pg/mL, 125pg/mL, 62.5pg/mL, and the last microcentrifuge tube with Standard Diluent is the blank as 0pg/mL.
3. Detection Reagent A and Detection Reagent B - Briefly spin or centrifuge the stock Detection A and Detection B before use. Dilute to the working concentration with Assay Diluent A and B, respectively (1:100).
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. 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 once 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.

Préparation de l'échantillon:

• 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).

Précision du teste: 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

Stockage

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

Stock: 4 °C/-20 °C

Stockage commentaire:

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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Date de péremption: 6 months

Références pour Hepcidin Kit ELISA (ABIN7059992)

Das, Sandhya, G, Kumar, Singh, Bal, Kumari, Mukhopadhyay: "Leishmania donovani inhibits ferroportin translation by modulating FBXL5-IRP2 axis for its growth within host macrophages." dans: Cellular microbiology, Vol. 20, Issue 7, pp. e12834, (2018) (PubMed).

Mao, Chen, Tang, Wang, Shu: "Hepcidin links gluco-toxicity to pancreatic beta cell dysfunction by inhibiting Pdx-1 expression." dans: Endocrine connections, Vol. 6, Issue 3, pp. 121-128, (2017) (PubMed).

Cheng, Zhou, Li, Liu, Wang, Zhang: "The effects of polysaccharides from the root of Angelica sinensis on tumor growth and iron metabolism in H22-bearing mice." dans: Food & function, Vol. 7, Issue 2, pp. 1033-9, (2016) (PubMed).

Gotardo, Caria, de Oliveira, Rocha, Ribeiro, Gambero: "Effects of iron supplementation in mice with hypoferremia induced by obesity." dans: Experimental biology and medicine (Maywood, N.J.), Vol. 241, Issue 18, pp. 2049-2055, (2016) (PubMed).

Wang, Tang, Xin, Zhu: "S-Propargyl-Cysteine, a Novel Hydrogen Sulfide Donor, Inhibits Inflammatory Hepcidin and Relieves Anemia of Inflammation by Inhibiting IL-6/STAT3 Pathway." dans: PLoS ONE, Vol. 11, Issue 9, pp. e0163289, (2016) (PubMed).

Gasperini, Meneghetti, Legname, Benetti: "In Absence of the Cellular Prion Protein, Alterations in Copper Metabolism and Copper-Dependent Oxidase Activity Affect Iron Distribution." dans: Frontiers in neuroscience, Vol. 10, pp. 437, (2016) (PubMed).

Shin, Chung, Joe, Pae, Chang, Cho, Ryter, Chung: "Pretreatment with CO-releasing molecules suppresses hepcidin expression during inflammation and endoplasmic reticulum stress through inhibition of the STAT3 and CREBH pathways." dans: Blood, Vol. 119, Issue 11, pp. 2523-32, (2012) (PubMed).

Détail du antigène

Antigène: Hepcidin (HAMP) (Hepcidin Antimicrobial Peptide (HAMP))

Autre désignation: Hepcidin (Hepc) (HAMP Produits)

Synonymes: HEPC Kit ELISA, HFE2B Kit ELISA, LEAP1 Kit ELISA, PLTR Kit ELISA, Hepcidin Kit ELISA, Hamp1 Kit ELISA, Hepc Kit ELISA, Hepc1 Kit ELISA, HAMP Kit ELISA, hepcidin Kit ELISA, hamp1 Kit ELISA, hep1 Kit ELISA, hepcidin antimicrobial peptide Kit ELISA, hepcidin-like Kit ELISA, hepcidin-1 Kit ELISA, Ham percentage Kit ELISA, HAMP Kit ELISA, Hamp Kit ELISA, hamp Kit ELISA, LOC100135935 Kit ELISA, LOC106584587 Kit ELISA

Pathways: Hormone Activity, Transition Metal Ion Homeostasis