Insulin Kit ELISA

N° du produit ABIN1979556

Détail du produit Insulin Kit ELISA

Antigène: Insulin (INS)

Reactivité: Rat

Méthode de détection: Colorimetric

Type de méthode: Sandwich ELISA

Gamme de detection: 5-300 μIU/mL

Seuil minimal de détection: 5 μIU/mL

Application: ELISA

Fonction: Rat Insulin ELISA Kit for cell culture supernatants, plasma, and serum samples.

Type d'échantillon: Serum, Plasma, Cell Culture Supernatant

Analytical Method: Quantitative

Specificité: This ELISA antibody pair detects rat, mouse, human, porcine and bovine Insulin and Proinsulin.

Sensibilité: 5 μIU/mL

Attributs du produit:

• Strip plates and additional reagents allow for use in multiple experiments
• Quantitative protein detection
• Establishes normal range
• The best products for confirmation of antibody array data

Ingrédients:

• Pre-Coated 96-well Strip Microplate
• Wash Buffer
• Stop Solution
• Assay Diluent(s)
• Lyophilized Standard
• Biotinylated Detection Antibody
• Streptavidin-Conjugated HRP
• TMB One-Step Substrate

Matériel non inclus:

• Distilled or deionized water
• Precision pipettes to deliver 2 μL to 1 μL volumes
• Adjustable 1-25 μL pipettes for reagent preparation
• 100 μL and 1 liter graduated cylinders
• Tubes to prepare standard and sample dilutions
• Absorbent paper
• Microplate reader capable of measuring absorbance at 450nm
• Log-log graph paper or computer and software for ELISA data analysis

Information d'application

Indications d'application: Recommended Dilution for serum and plasma samples2 fold

Volume d'échantillon: 100 μL

Plaque: Pre-coated

Protocole:

1. Prepare all reagents, samples and standards as instructed in the manual.
2. Add 100 μL of standard or sample to each well.
3. Incubate 2.5 h at RT or O/N at 4 °C.
4. Add 100 μL of prepared biotin antibody to each well.
5. Incubate 1 h at RT.
6. Add 100 μL of prepared Streptavidin solution to each well.
7. Incubate 45 min at RT.
8. Add 100 μL of TMB One-Step Substrate Reagent to each well.
9. Incubate 30 min at RT.
10. Add 50 μL of Stop Solution to each well.
11. Read at 450 nm immediately.

Préparation des réactifs:

1. Bring all reagents and samples to room temperature (18 - 25 °C) before use. 2. Sample dilution: If your samples need to be diluted, Assay Diluent C (Item L) should be used for dilution of serum/plasma/culture supernatants/urine. Suggested dilution for normal serum/plasma: 2 fold. The Rat Insulin ELISA Kit Protocol 3 Please note that levels of the target protein may vary between different specimens. Optimal dilution factors for each sample must be determined by the investigator. 3. Assay Diluent B (Item E) should be diluted 5-fold with deionized or distilled water before use. 4. Preparation of standard: Briefly spin the vial of Item C. Add 400 µL Assay Diluent C (Item L) into Item C vial to prepare a 1,400 µIU/ml standard solution. Dissolve the powder thoroughly by a gentle mix. Add 150 µL Insulin standard (1,400 µIU/ml) from the vial of Item C, into a tube with 550 µL Assay Diluent C to prepare a 300 µIU/ml standard solution. Pipette 250myl Assay Diluent C into each tube. Use the 300 µIU/ml standard solution to produce a dilution series (shown below). Mix each tube thoroughly before the next transfer. Assay Diluent C serves as the zero standard (0 µIU/ml). 250 µL 250 µL 250 µL 250 µL 250myl 180 µL Standard + 450 µL 250 µL 300 150 75 37.5 18.75 9.38 4.69 0 µIU/ml µIU/ml µIU/ml µIU/ml µIU/ml µIU/ml µIU/ml µIU/ml 5. If the Wash Concentrate (20x) (Item B) contains visible crystals, warm to room temperature and mix gently until dissolved. Dilute 20 mL of Wash Buffer Concentrate into deionized or distilled water to yield 400 mL of 1x Wash Buffer. The Rat Insulin ELISA Kit Protocol 4 6. Briefly spin the Detection Antibody vial (Item F) before use. Add 100 µL of 1x Assay Diluent B (Item E) into the vial to prepare a detection antibody concentrate. Pipette up and down to mix gently (the concentrate can be stored at 4 °C for 5 days). The detection antibody concentrate should be diluted 80-fold with 1x Assay Diluent B and used in step 4 of Part VI Assay Procedure. 7. Briefly spin the HRP-Streptavidin concentrate vial (Item G) and pipette up and down to mix gently before use. HRP-Streptavidin concentrate should be diluted 400-fold with 1x Assay Diluent B (Item E). For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 30 µL of HRP-Streptavidin concentrate into a tube with 12 mL 1x Assay Diluent B to prepare a 400-fold diluted HRP-Streptavidin solution (don't store the diluted solution for next day use). Mix well.

Procédure de l'essai:

1. Bring all reagents and samples to room temperature (18 - 25 °C) before use. It is recommended that all standards and samples be run at least in duplicate. 2. Add 100 µL of each standard (see Reagent Preparation step 2) and sample into appropriate wells. Cover well and incubate for 2.5 hours at room temperature or over night at 4 °C with gentle shaking. 3. Discard remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels. 4. Add 100 µL of 1x prepared biotinylated antibody (Reagent Preparation step 6) to each well. Incubate for 1 hour at room temperature with gentle shaking. 5. Discard the solution. Repeat the wash as in step 3. 6. Add 100 µL of prepared Streptavidin solution (see Reagent Preparation step 7) to each well. Incubate for 45 minutes at room temperature with gentle shaking. 7. Discard the solution. Repeat the wash as in step 3. 8. Add 100 µL of TMB One-Step Substrate Reagent (Item H) to each well. Incubate for 30 minutes at room temperature in the dark with gentle shaking. 9. Add 50 µL of Stop Solution (Item I) to each well. Read at 450 nm immediately.

Calcul des résultats:

Calculate the mean absorbance for each set of duplicate standards, controls and samples, and subtract the average zero standard optical density. Plot the standard curve on log-log graph paper or using Sigma plot software, with standard concentration on the x-axis and absorbance on the y-axis. Draw the best-fit straight line through the standard points.

Précision du teste: Intra-Assay: CV<10%
Inter-Assay: CV<12%

Restrictions: For Research Use only

Stockage

Conseil sur la manipulation: Avoid repeated freeze-thaw cycles.

Stock: -20 °C

Stockage commentaire: The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.

Date de péremption: 6 months

Références pour Insulin Kit ELISA (ABIN1979556)

Ding, Tong, Feng, Chen, Wang, Wang, Li, Zhu, Zhou: "Adipose afferent reflex response to insulin is mediated by melanocortin 4 type receptors in the paraventricular nucleus in insulin resistance rats." dans: Acta physiologica (Oxford, England), Vol. 214, Issue 4, pp. 450-66, (2015) (PubMed).

Shihabudeen, Roy, James, Thirumurugan: "Chenodeoxycholic acid, an endogenous FXR ligand alters adipokines and reverses insulin resistance." dans: Molecular and cellular endocrinology, Vol. 414, pp. 19-28, (2015) (PubMed).

Lee, Seo, Ryu, Kwon: "The effect of swimming exercise and powdered-Salicornia herbacea L. ingestion on glucose metabolism in STZ-induced diabetic rats." dans: Journal of exercise nutrition & biochemistry, Vol. 19, Issue 3, pp. 235-45, (2015) (PubMed).

Al-Gayyar, Alyoussef, Hamdan, Abbas, Darweish, El-Hawwary: "Cod liver oil ameliorates sodium nitrite-induced insulin resistance and degradation of rat hepatic glycogen through inhibition of cAMP/PKA pathway." dans: Life sciences, Vol. 120, pp. 13-21, (2014) (PubMed).

Defranoux, Jutisz, Jerome, Job: "[Study of the effects of human growth hormone in a patient with pituitary dwarfism]." dans: Annales d'endocrinologie, Vol. 26, Issue 4, pp. 501-12 (PubMed).

Détail du antigène

Antigène: Insulin (INS)

Autre désignation: Insulin (INS Produits)

Synonymes: IDDM2 Kit ELISA, ILPR Kit ELISA, IRDN Kit ELISA, MODY10 Kit ELISA, ins1 Kit ELISA, xins Kit ELISA, ins1-a Kit ELISA, Insulin Kit ELISA, AA986540 Kit ELISA, Ins-2 Kit ELISA, InsII Kit ELISA, Mody Kit ELISA, Mody4 Kit ELISA, proinsulin Kit ELISA, zgc:109842 Kit ELISA, igf2-A Kit ELISA, ins Kit ELISA, ins-a Kit ELISA, ins-b Kit ELISA, insulin Kit ELISA, insulin precursor Kit ELISA, insulin II Kit ELISA, preproinsulin Kit ELISA, insulin L homeolog Kit ELISA, insulin S homeolog Kit ELISA, INS Kit ELISA, INS-IGF2 Kit ELISA, ins Kit ELISA, Ins Kit ELISA, PIN Kit ELISA, Ins2 Kit ELISA, ins.L Kit ELISA, ins.S Kit ELISA

ID gène: 24506

UniProt: P01323

Pathways: Signalisation NF-kappaB, Signalisation RTK, Positive Regulation of Peptide Hormone Secretion, Peptide Hormone Metabolism, Hormone Activity, Carbohydrate Homeostasis, ER-Nucleus Signaling, Regulation of Carbohydrate Metabolic Process, Feeding Behaviour, Autophagy, Negative Regulation of intrinsic apoptotic Signaling, Brown Fat Cell Differentiation, Positive Regulation of fat Cell Differentiation