Insulin ELISA Kit

Catalog No. ABIN6974595

Product Details Insulin ELISA Kit

Target: Insulin (INS)

Reactivity: Rat

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 15.6 nlU/mL - 1000 nlU/mL

Minimum Detection Limit: 15.6 nlU/mL

Application: ELISA

Purpose: For the quantitative determination of rat insulin (INS) concentrations in serum, plasma, cell culture supernates.

Sample Type: Cell Culture Supernatant, Plasma, Serum

Analytical Method: Quantitative

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

Sensitivity: 3.9 nlU/mL

Components:

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

Application Details

Application Notes: Optimal working dilution should be determined by the investigator.

Sample Volume: 100 μL

Assay Time: 1 - 4.5 h

Plate: Pre-coated

Protocol:

1. Prepare reagents, samples and standards as instructed.
2. Add 100 µL standard or sample to each well. Incubate 2 hours at 37 °C.
3. Remove the liquid of each well, don't wash.
4. Add 100 µL Biotin-antibody (1x) to each well. Incubate 1 hour at 37 °C.
5. Aspirate and wash 3 times.
6. Add 100 µL HRP-avidin (1x) to each well. Incubate 1 hour at 37 °C
7. Aspirate and wash 5 times.
8. Add 90 μL of TMB Substrate to each well. Incubate for 15-30 minutes at 37 °C. Protect from light.
9. Add 50 µL Stop Solution to each well. Read at 450 nm within 5 minutes.

Reagent Preparation:

1. Biotin-antibody (1x) - Centrifuge the vial before opening. Biotin-antibody requires a 100-fold dilution. A suggested 100-fold dilution is 10 μL of Biotin-antibody + 990 μL of Biotin-antibody Diluent.
2. HRP-avidin (1x) - Centrifuge the vial before opening. HRP-avidin requires a 100-fold dilution. A suggested 100-fold dilution is 10 μL of HRP-avidin + 990 μL of HRP-avidin Diluent.
3. 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).
4. 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 1000 nIU/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 250 μL of Sample Diluent into each tube (S0-S6). Use the stock solution to produce a 2-fold dilution series (below). Mix each tube thoroughly before the next transfer. The undiluted Standard serves as the high standard (1000 nIU/mL). Sample Diluent serves as the zero standard (0 nIU/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.

Sample Preparation:

• 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).

Note:

Recommend to dilute the serum or plasma samples with Sample Diluent (1:200) before test. The suggested 200-fold dilution can be achieved by adding 5 μL sample to 45 μL of Sample Diluent. Complete the 200-fold dilution by adding15 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.

Assay Precision: 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

Handling

Storage: 4 °C,-20 °C

Storage Comment: 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 Biotin-antibody don't make recent use, better keep it store at HRP-avidin -20°C. Biotin-antibody Diluent Opened kit HRP-avidin Diluent Sample May be stored for up to 1 month at 2 - 8°C. Diluent Wash Buffer TMB Substrate Stop Solution *Provided this is within the expiration date of the kit.

Expiry Date: 6 months

References for Insulin ELISA Kit (ABIN6974595)

Chen, Xu, Wu, Han, Yu, Ge, Chen: "Impaired learning and memory in rats induced by a high-fat diet: Involvement with the imbalance of nesfatin-1 abundance and copine 6 expression." in: Journal of neuroendocrinology, Vol. 29, Issue 4, (2018) (PubMed).

El-Shemi, Kensara, Alsaegh, Mukhtar et al.: "Pharmacotherapy with Thymoquinone Improved Pancreatic β-Cell Integrity and Functional Activity, Enhanced Islets Revascularization, and Alleviated Metabolic and Hepato-Renal Disturbances in ..." in: Pharmacology, Vol. 101, Issue 1-2, pp. 9-21, (2018) (PubMed).

Elseweidy, Amin, Atteia, Aly: "Nigella sativa Oil and Chromium Picolinate Ameliorate Fructose-Induced Hyperinsulinemia by Enhancing Insulin Signaling and Suppressing Insulin-Degrading Enzyme in Male Rats." in: Biological trace element research, Vol. 184, Issue 1, pp. 119-126, (2018) (PubMed).

Hou, Hu, Yang, Chen: "Antihypertensive effects of Tartary buckwheat flavonoids by improvement of vascular insulin sensitivity in spontaneously hypertensive rats." in: Food & function, Vol. 8, Issue 11, pp. 4217-4228, (2018) (PubMed).

Zhao, Zhang, Ma, Tian, Shen, Zhou: "A combination of quercetin and resveratrol reduces obesity in high-fat diet-fed rats by modulation of gut microbiota." in: Food & function, Vol. 8, Issue 12, pp. 4644-4656, (2018) (PubMed).

Zheng, Niu: "Leptin-induced basal Akt phosphorylation and its implication in exercise-mediated improvement of insulin sensitivity." in: Biochemical and biophysical research communications, Vol. 496, Issue 1, pp. 37-43, (2018) (PubMed).

Li, Ni, Zhao, Liu, Lai, Di, Xie, Song, Wang, Zhang, Liu: "Melatonin attenuates smoking-induced hyperglycemia via preserving insulin secretion and hepatic glycogen synthesis in rats." in: Journal of pineal research, Vol. 64, Issue 4, pp. e12475, (2018) (PubMed).

Long, Zhang, Sun, Liu, Liao, Wu, Wang, Hai: "Evolution of metabolic disorder in rats fed high sucrose or high fat diet: Focus on redox state and mitochondrial function." in: General and comparative endocrinology, Vol. 242, pp. 92-100, (2017) (PubMed).

Zhang, Yu, Xu, Wang, Ji, Gu, Yang, Zhu, Dong, Wang: "High-fat diet aggravates 2,2',4,4'-tetrabromodiphenyl ether-inhibited testosterone production via DAX-1 in Leydig cells in rats." in: Toxicology and applied pharmacology, Vol. 323, pp. 1-8, (2017) (PubMed).

Shen, Yang, Yan, Zheng, Liang, Cai, Liao: "Fetuin A promotes lipotoxicity in ? cells through the TLR4 signaling pathway and the role of pioglitazone in anti-lipotoxicity." in: Molecular and cellular endocrinology, Vol. 412, pp. 1-11, (2015) (PubMed).

Jia, Ma, Liu, Zhou, He, Xu, Ren, Xu, Tian: "Metformin prevents DMH-induced colorectal cancer in diabetic rats by reversing the warburg effect." in: Cancer medicine, (2015) (PubMed).

Wu, Sun, Yin, Xu, Wang, Lin, Lin, Lin: "Different effect of handle region peptide on β-cell function in different sexes of rats neonatally treated with sodium L-glutamate." in: Medical science monitor basic research, Vol. 21, pp. 33-40, (2015) (PubMed).

Secher, Østergaard, Iversen, Lambertsen, Clausen, Tønnesen, Granfeldt: "Preserved Cerebral Microcirculation After Cardiac Arrest in a Rat Model." in: Microcirculation (New York, N.Y. : 1994), Vol. 22, Issue 6, pp. 464-74, (2015) (PubMed).

Perimenis, Bouckenooghe, Delplanque, Moitrot, Eury, Lobbens, Gosset, Devisme, Duvillie, Abderrahmani, Storme, Fontaine, Froguel, Vambergue: "Placental antiangiogenic prolactin fragments are increased in human and rat maternal diabetes." in: Biochimica et biophysica acta, Vol. 1842, Issue 9, pp. 1783-93, (2014) (PubMed).

Yin, Lin, Xu, Sun, Lin, Lin: "Handle Region Peptide Ameliorating Insulin Resistance but Not β Cell Functions in Male Rats Neonatally Treated with Sodium L-Glutamate." in: International journal of endocrinology, Vol. 2013, pp. 493828, (2014) (PubMed).

Zeng, Wang, Li, Shen, Wang, Yu, Wang: "AKAP150 mobilizes cPKC-dependent cardiac glucotoxicity." in: American journal of physiology. Endocrinology and metabolism, Vol. 307, Issue 4, pp. E384-97, (2014) (PubMed).

Lu, Zhang, Zheng, Jiang, Chen: "Branched-chain amino acids supplementation protects streptozotocin-induced insulin secretion and the correlated mechanism." in: BioFactors (Oxford, England), (2014) (PubMed).

Wang, Luo, Wang, Li, Wang, Sun, Zhang, Su, Ma, Zeng, Wang, Ren, Cao: "Glucagon-like peptide-1 protects against cardiac microvascular injury in diabetes via a cAMP/PKA/Rho-dependent mechanism." in: Diabetes, Vol. 62, Issue 5, pp. 1697-708, (2013) (PubMed).

Zhang, Sun, Xiao, Zhou, Wang, Gu, Qiu, Zhang, Xu, Zhen, Wang, Wang: "Mechanism of BDE209-induced impaired glucose homeostasis based on gene microarray analysis of adult rat liver." in: Archives of toxicology, Vol. 87, Issue 8, pp. 1557-67, (2013) (PubMed).

Wang, Liu, Zhang, Zhang, Liao, Wang, Li, Qin, Hai: "Oleanolic acid improves hepatic insulin resistance via antioxidant, hypolipidemic and anti-inflammatory effects." in: Molecular and cellular endocrinology, Vol. 376, Issue 1-2, pp. 70-80, (2013) (PubMed).

Target Details for Insulin

Target: Insulin (INS)

Alternative Name: Insulin,INS (INS Products)

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

Background:

Abbreviation: INS

Alias: N/A

UniProt: P01322

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