Insulin CLIA Kit

Catalog No. ABIN504795

The Human Insulin ELISA Kit (ABIN504795) is a Chemiluminescent ELISA Kit designed to quantify Human Insulin.

Quick Overview for Insulin CLIA Kit (ABIN504795)

Target: Insulin (INS)

Reactivity: Human

Detection Method: Chemiluminescent

Method Type: Sandwich ELISA

Application: ELISA

Product Details Insulin CLIA Kit

Purpose: Human insulin is a peptide produced in the beta cells of the pancreas and is responsible for the metabolism and storage of carbohydrates. As a result of biofeedback the insulin levels increase with intake of sugars and decline when sugar content is low for absorption. In the diabetic population the mechanism of insulin production is impaired because of genetic predispositions (Type I) or because of lifestyle and/or hereditary factors (Type II). In such cases either the insulin production has to be boosted by medication or it has to be supplemented by oral or intravenous methods. The quantitative determination of insulin can help in dose selection the patient has to be subjected to. On the other hand the circulatory insulin can be found at much higher levels in patients with pancreatic tumors. These tumors secrete abnormally high levels of insulin and thus cause hypoglycemia. Accordingly, fasting hypoglycemia associated with inappropriately high concentrations of insulin strongly suggests an islet-cell tumor (insulinoma). To distinguish insulinomas from factitious hypoglycemia due to insulin administration, serum C-peptide values are recommended. These insulinomas can be localized by provocative intravenous doses of tolbutamide and calcium.

Analytical Method: Quantitative

Characteristics: The Quantitative Determination of Insulin Concentration in Human Serum by a Microplate Chemiluminescence Assay

Application Details

Application Notes: All products that contain human serum have been found to be non-reactive for Hepatitis B Surface Antigen, HIV 1&2 and HCV Antibodies by FDA required tests. Since no known test can offer complete assurance that infectious agents are absent, all human serum products should be handled as potentially hazardous and capable of transmitting disease. Good laboratory procedures for handling blood products can be found in the Center for Disease Control / National Institute of Health, "Biosafety in Microbiological and Biomedical Laboratories," 2nd Edition, 1988, HHS Publication No. (CDC) 88-8395.

Sample Volume: 50 μL

Plate: Pre-coated

Protocol:

Specimien Collection and Preparation:

The specimens shall be blood, serum or plasma in type and the usual precautions in the collection of venipuncture samples should be observed. For accurate comparison to established normal values, a fasting morning serum sample should be obtained. The blood should be collected in a plain redtop venipuncture tube without additives or anti-coagulants (for serum) or evacuated tube(s) containing EDTA or heparin.. Allow the blood to clot for serum samples. Centrifuge the specimen to separate the serum or plasma from the cells. Samples may be refrigerated at 2_x001E_8oC for a maximum period of five (5) days. If the specimen(s) cannot be assayed within this time, the sample(s) may be stored at temperatures of _x001E_20oC for up to 30 days. Avoid repetitive freezing and thawing. When assayed in duplicate, 0.100ml of the specimen is required

Reagent Preparation:

1. Wash Buffer Dilute contents of Wash Concentrate to 1000ml with distilled or deionized water in a suitable storage container. Store at room temperature (20-27(C) for up to 60 days. 2. Working Signal Reagent Solution - Store at 2 - 8(C. Determine the amount of reagent needed and prepare by mixing equal portions of Signal Reagent A and Signal Reagent B in a clean container. For example, add 1 ml of A and 1ml of B per two (2) eight well strips (A slight excess of solution is made). Discard the unused portion if not used within 36 hours after mixing. If complete utilization of the reagents is anticipated, within the above time constraint, pour the contents of Signal Reagent B into Signal Reagent A and label accordingly. _x000E_

Test Procedure:

Before proceeding with the assay, bring all reagents, serum references and controls to room temperature (20 - 27( C). 1. Format the microplates wells for calibrator, control and patient specimen to be assayed in duplicate. Replace any unused microwell strips back into the aluminum bag, seal and store at 2-8(C. 2. Pipette 0.050 ml (50l) of the appropriate calibrators, controls and samples into the assigned wells. 3. Add 0.100 ml (100l) of the Insulin Tracer Reagent to each well. It is very important to dispense all reagents close to the bottom of the microwell. 4. Swirl the microplate gently for 20-30 seconds to mix. Cover with a plastic wrap. 5. Incubate for 60 minutes at room temperature 6. Discard the contents of the microplate by decantation or aspiration. If decanting, tap and blot the plate dry with absorbent paper. 7. Add 350l of wash buffer (see Reagent Preparation Section), decant (tap and blot) or aspirate. Repeat four (4) additional times for a total of five (5) washes. An automatic or manual plate washer can be used. Follow the manufacturers instruction for proper usage. If a squeeze bottle is used, fill each well to the top by squeezing the container. Avoiding air bubbles. Decant the wash and repeat four (4) additional times. Add 0.100 ml (100l) of working signal reagent solution to all wells (see Reagent Preparation Section). Always add reagents in the same order to minimize reaction time differences between wells. Incubate at room temperature for five (5) minutes in the dark. 10. Read the RLUs (Relative Light Units) using a 96 well microplate luminometer for 0.2 1.0 seconds per well. The results should be read within thirty (30) minutes of adding the stop solution.

Restrictions: For Research Use only

Handling

Target Details

Target: Insulin (INS)

Alternative Name: Insulin

Background: Human insulin is a peptide produced in the beta cells of the pancreas and is responsible for the metabolism and storage of carbohydrates. As a result of biofeedback the insulin levels increase with intake of sugars and decline when sugar content is low for absorption. In the diabetic population the mechanism of insulin production is impaired because of genetic predispositions (Type I) or because of lifestyle and/or hereditary factors (Type II). In such cases either the insulin production has to be boosted by medication or it has to be supplemented by oral or intravenous methods. The quantitative determination of insulin can help in dose selection the patient has to be subjected to. On the other hand the circulatory insulin can be found at much higher levels in patients with pancreatic tumors. These tumors secrete abnormally high levels of insulin and thus cause hypoglycemia. Accordingly, fasting hypoglycemia associated with inappropriately high concentrations of insulin strongly suggests an islet-cell tumor (insulinoma). To distinguish insulinomas from factitious hypoglycemia due to insulin administration, serum C-peptide values are recommended. (Please see Monobind C-Peptide Microwell CLIA Cat#2775-300). These insulinomas can be localized by provocative intravenous doses of tolbutamide and calcium.

Pathways: NF-kappaB Signaling, RTK Signaling, 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