APOA5 Kit ELISA

N° du produit ABIN6973943

Détail du produit APOA5 Kit ELISA

Antigène: APOA5 (Apolipoprotein A-V (APOA5))

Reactivité: Humain

Méthode de détection: Colorimetric

Type de méthode: Competition ELISA

Gamme de detection: 2.5 ng/mL - 40 ng/mL

Seuil minimal de détection: 2.5 ng/mL

Application: ELISA

Fonction: For the quantitative determination of human apolipoprotein A5 (Apo-A5) concentrations in serum, plasma, tissue homogenates.

Type d'échantillon: Plasma, Serum, Tissue Homogenate

Analytical Method: Quantitative

Specificité: This assay has high sensitivity and excellent specificity for detection of human Apo-A5. No significant cross-reactivity or interference between human Apo-A5 and analogues was observed. Note: Limited by current skills and knowledge, it is impossible for us to complete the cross-reactivity detection between human Apo-A5 and all the analogues, therefore, cross reaction may still exist.

Sensibilité: 1.25 ng/mL

Ingrédients:

• Assay plate
• Standard
• HRP-conjugate (100 x concentrate)
• Sample Diluent
• HRP-conjugate Diluent
• Wash Buffer (25 x concentrate)
• TMB Substrate
• Stop Solution
• Adhesive Strip

Stop Solution

Adhesive Strip

Information d'application

Indications d'application: Optimal working dilution should be determined by the investigator.

Volume d'échantillon: 50 μL

Durée du test: 1 - 4.5 h

Plaque: Pre-coated

Protocole:

1. Prepare reagents, samples and standards as instructed.
2. Set a Blank well without any solution.
3. Add 50 µL standard or sample to each well.
4. Add 50 µL HRP-conjugate (1x) to each well (Not to Blank well).
5. Incubate 1 hour at 37 °C
6. Aspirate and wash 5 times.
7. Add 90 μL of TMB Substrate to each well. Incubate for 20 minutes at 37 °C. Protect from light.
8. Add 50 µL Stop Solution to each well. Read at 450 nm within 5 minutes.

Préparation des réactifs:

1. HRP-conjugate (1x) - Centrifuge the vial before opening. HRP-conjugate requires a 100-fold dilution. A suggested 100-fold dilution is 10 μL of HRP-conjugate + 990 μL of HRP-conjugate Diluent.
2. Wash Buffer(1x)- If crystals have formed in the concentrate, warm up to room temperature and mix gently until the crystals have completely dissolved. Dilute 20 mL of Wash Buffer Concentrate (25 x) into deionized or distilled water to prepare 500 mL of Wash Buffer (1 x).
3. Standard Centrifuge the standard vial at 6000-10000rpm for 30s. Reconstitute the Standard with 1.0 mL of Sample Diluent. Do not substitute other diluents. This reconstitution produces a stock solution of 40 ng/mL. Mix the standard to ensure complete reconstitution and allow the standard to sit for a minimum of 15 minutes with gentle agitation prior to making dilutions. Pipette 150 μl of Sample Diluent into each tube (S0-S4). Use the stock solution to produce a 2-fold dilution series. Mix each tube thoroughly before the next transfer. The undiluted Standard serves as the high standard (40 ng/ml). Sample Diluent serves as the zero standard (0 ng/ml).

Note:

• Kindly use graduated containers to prepare the reagent. Please don't prepare the reagent directly in the Diluent vials provided in the kit.
• Bring all reagents to room temperature (18-25 °C) before use for 30 min.
• Prepare fresh standard for each assay. Use within 4 hours and discard after use.
• Making serial dilution in the wells directly is not permitted.
• Please carefully reconstitute Standards according to the instruction, and avoid foaming and mix gently until the crystals have completely dissolved. To minimize imprecision caused by pipetting, use small volumes and ensure that pipettors are calibrated. It is recommended to suck more than 10 μL for once pipetting.
• Distilled water is recommended to be used to make the preparation for reagents. 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 denaturation may occur in these samples, leading to false results. Samples should therefore be stored for a short period at 2 - 8 °C or aliquoted at -20 °C (≤1 month) or -80 °C (≤ 3 months). Repeated freeze-thaw cycles should be avoided. Prior to assay, the frozen samples should be slowly thawed and centrifuged to remove precipitates.
• If the sample type is not specified in the instructions, a preliminary test is necessary to determine compatibility with the kit.
• If a lysis buffer is used to prepare tissue homogenates or cell culture supernatant, there is a possibility of 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 the range of the standard curve, the optimal sample dilution for the particular experiment has to be determined.

Note:

Recommend to dilute the serum or plasma samples with Sample Diluent(1:100) before test. The suggested 100-fold dilution can be achieved by adding 10 μL sample to 40 μL of Sample Diluent. Complete the 100-fold dilution by adding 15 μL of this solution to 285 μL of Sample Diluent. The recommended dilution factor is for reference only. The optimal dilution factor should be determined by users according to their particular experiments. 6

Précision du teste: Intra-assay Precision (Precision within an assay): CV%<8% Three samples of known concentration were tested twenty times on one plate to assess.
Inter-assay Precision (Precision between assays): CV%<10% Three samples of known concentration were tested in twenty assays to assess.

Restrictions: For Research Use only

Stockage

Stock: 4 °C,-20 °C

Stockage commentaire: Unopened kit Store at 2 - 8°C. Do not use the kit beyond the expiration date. May be stored for up to 1 month at 2 - 8°C. Coated assay Try to keep it in a sealed aluminum foil bag, plate and avoid the damp. Standard May be stored for up to 1 month at 2 - 8° C. If don't make recent use, better keep it store at HRP-conjugate -20°C. Opened kit HRP-conjugate Diluent Sample Diluent May be stored for up to 1 month at 2 - 8°C. Wash Buffer TMB Substrate Stop Solution *Provided this is within the expiration date of the kit.

Date de péremption: 6 months

Références pour APOA5 Kit ELISA (ABIN6973943)

Lightbourne, Wolska, Abel, Rother, Walter, Kushchayeva, Auh, Shamburek, Remaley, Muniyappa, Brown: "Apolipoprotein CIII and Angiopoietin-like Protein 8 are Elevated in Lipodystrophy and Decrease after Metreleptin." dans: Journal of the Endocrine Society, Vol. 5, Issue 2, pp. bvaa191, (2021) (PubMed).

Zhu, Zhang, Zhou, Li, Li, Fang, Yang, Shan, Li, Chen, Zhou, Ye, Yu, Li, Cai, Liu, Zhang, Wang, Lai, Ruan, Sun, Zhang, Wang, Liu, Xu, Ling, Xu, Zhang, Lv, Yuan, Zhang, Zhang, Shi, Lai: "Susceptibility loci for metabolic syndrome and metabolic components identified in Han Chinese: a multi-stage genome-wide association study." dans: Journal of cellular and molecular medicine, Vol. 21, Issue 6, pp. 1106-1116, (2018) (PubMed).

Kim, Kim, Yoo, Lee, Chae, Lee, Lee: "A promoter variant of the APOA5 gene increases atherogenic LDL levels and arterial stiffness in hypertriglyceridemic patients." dans: PLoS ONE, Vol. 12, Issue 12, pp. e0186693, (2018) (PubMed).

Ngaosuwan, Houngngam, Limpisook, Plengpanich, Khovidhunkit: "Apolipoprotein A-V is not a major determinant of triglyceride levels during human sepsis." dans: Journal of critical care, Vol. 30, Issue 4, pp. 727-31, (2015) (PubMed).

Détail du antigène

Antigène: APOA5 (Apolipoprotein A-V (APOA5))

Autre désignation: apolipoprotein A-V (APOA5 Produits)

Synonymes: APOAV Kit ELISA, RAP3 Kit ELISA, 1300007O05Rik Kit ELISA, Apoav Kit ELISA, apo-AV Kit ELISA, apoA-V Kit ELISA, APOA5 Kit ELISA, MGC140487 Kit ELISA, rap3 Kit ELISA, apoav Kit ELISA, apoa-v Kit ELISA, apolipoprotein A5 Kit ELISA, apolipoprotein A-V Kit ELISA, apolipoprotein A5 L homeolog Kit ELISA, APOA5 Kit ELISA, Apoa5 Kit ELISA, apoa5.L Kit ELISA, apoa5 Kit ELISA

Sujet:

Abbreviation: APOA5

Alias: APOAV, FLJ97995, MGC126836, MGC126838, RAP3, apolipoprotein A5|regeneration-associated protein 3

UniProt: Q6Q788

Pathways: Regulation of Lipid Metabolism by PPARalpha, Lipid Metabolism