Insulin ELISA Kit

Catalog No. ABIN7054819

Product Details Insulin ELISA Kit

Target: Insulin (INS)

Reactivity: Mouse

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 3.13 pg/mL - 200 pg/mL

Minimum Detection Limit: 3.13 pg/mL

Application: ELISA

Purpose: The kit is a sandwich enzyme immunoassay technique for the in vitro quantitative measurement in various sample types.

Sample Type: Cell Culture Supernatant, Plasma, Serum

Analytical Method: Quantitative

Specificity: This kit recognizes Mouse INS in samples. No Significant cross-reactivity or interference between Mouse INS and analogues was observed.

Sensitivity: 1.88 pg/mL

Components:

• Pre-coated, ready to use 96-well strip plate, flat buttom
• Plate sealer for 96 wells
• Reference Standard
• Reference Standard & Sample Diluent
• Biotinylated Detection Antibody (100 x concentrate)
• HRP Conjugate (100 x concentrate)
• Biotinylated Detection Antibody Diluent
• HRP Conjugate Diluent
• Substrate Reagent
• Stop Solution
• Wash Buffer (25 x concentrate)
• Instruction manual

Application Details

Sample Volume: 100 μL

Assay Time: 3.5 h

Plate: Pre-coated

Protocol:

1. Add 100 µL standard or sample to each well. Incubate for 90 min at 37 °C.
2. Remove the liquid. Add 100 µL Biotinylated Detection Antibody. Incubate for 1 hour at 37 °C.
3. Aspirate and wash 3 times.
4. Add 100 µL HRP Conjugate. Incubate for 30 min at 37 °C.
5. Aspirate and wash 5 times.
6. Add 90 µL Substrate Reagent. Incubate for 15 min at 37 °C.
7. Add 50 µL Stop Solution. Read at 450 nm immediately.
8. Calculation of results.

Reagent Preparation:

1. Bring all reagents to room temperature (18~25 °C) before use. Follow the Microplate reader manual for set-up and preheat it for 15 min before OD measurement.
2. Wash Buffer: Dilute 30 mL of Concentrated Wash Buffer with 720 mL of deionized or distilled water to prepare 750 mL of Wash Buffer.Note: if crystals have formed in the concentrate, warm it in a 40 °C water bath and mix it gently until the crystals have completely dissolved
3. Standard working solution: Centrifuge the standard at 10,000xg for 1 min. Add 1.0 mL of Reference Standard &Sample Diluent, let it stand for 10 min and invert it gently several times. After it dissolves fully, mix it thoroughly with a pipette. This reconstitution produces a working solution of 200 pg/mL. Then make serial dilutions as needed. The recommended dilution gradient is as follows: 200, 100, 50, 25, 12.5, 6.25, 3.13, 0 pg/mL. Dilution method: Take 7 EP tubes, add 500 μLof Reference Standard & Sample Diluent to each tube. Pipette 500 μLof the 200 pg/mL working solution to the first tube and mix up to produce a 100 pg/mL working solution. Pipette 500 μLof the solution from the former tube into the latter one according to these steps. The illustration below is for reference. Note: the last tube is regarded as a blank. Don't pipette solution into it from the former tube.
4. Biotinylated Detection Antibody working solution: Calculate the required amount before the experiment (100 μL/well). In preparation, slightly more than calculated should be prepared. Centrifuge the stock tube before use, dilute the 100x Concentrated Biotinylated Detection Antibody to 1xworking solution with Biotinylated Detection Antibody Diluent.
5. Concentrated HRP Conjugate working solution: Calculate the required amount before the experiment (100 μL/well). In preparation, slightly more than calculated should be prepared. Dilute the 100x Concentrated HRP Conjugate to 1x working solution with Concentrated HRP Conjugate Diluent.

Sample Preparation:

• 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.

Assay Precision: Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Mouse INS were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Mouse INS were tested on 3 different plates, 20 replicates in each plate.
Both intra-CV and inter-CV are < 10 %.

Restrictions: For Research Use only

Handling

Storage: 4 °C,-20 °C

Storage Comment:

1. For unopened kit: All reagents should be stored according to the labels on the vials, so they are stable up to 12 months after receipt of the kit. The Reference Standard, Biotinylated Detection Antibody, HRP Conjugate and the 96-well stripe plate should be stored at -20 °C upon receipt while the other reagents should be stored at 4 °C.
2. For used kit: When the kit is used, the remaining reagents need to be stored according to the above storage condition. Besides, please return the unused wells to the foil pouch containing the desiccant pack, and zip-seal the foil pouch.

Expiry Date: 12 months

References for Insulin ELISA Kit (ABIN7054819)

Sun, Zhao, Liu, Sun, Zhang, Wang, Xu, Zhang, Wang: "Modulation of Gut Microbiota by Fucoxanthin During Alleviation of Obesity in High-Fat Diet-Fed Mice." in: Journal of agricultural and food chemistry, Vol. 68, Issue 18, pp. 5118-5128, (2021) (PubMed).

Singh, Bansal, Sodhi, Khare, Bishnoi, Kondepudi, Medhi, Kuhad: "Role of TRPV1/TRPV3 channels in olanzapine-induced metabolic alteration: Possible involvement in hypothalamic energy-sensing, appetite regulation, inflammation and mesolimbic pathway." in: Toxicology and applied pharmacology, Vol. 402, pp. 115124, (2021) (PubMed).

Hu, Chen, Qiu, Yang, Liu, Zhang, Zhang, Wang: "Intra-Pancreatic Insulin Nourishes Cancer Cells: Do Insulin-Receptor Antagonists such as PGG and EGCG Play a Role?" in: The American journal of Chinese medicine, Vol. 48, Issue 4, pp. 1005-1019, (2020) (PubMed).

Su, Mo, Ni, Ke, Bao, Xie, Xu, Xie, Chen: "Andrographolide Exerts Antihyperglycemic Effect through Strengthening Intestinal Barrier Function and Increasing Microbial Composition of Akkermansia muciniphila." in: Oxidative medicine and cellular longevity, Vol. 2020, pp. 6538930, (2020) (PubMed).

Singh, Bansal, Sodhi, Saroj, Medhi, Kuhad: "Modeling of antipsychotic-induced metabolic alterations in mice: An experimental approach precluding psychosis as a predisposing factor." in: Toxicology and applied pharmacology, Vol. 378, pp. 114643, (2020) (PubMed).

Yang, Chen, Duan, Ma, Liu, Li, Yang, Li, Zhao, Wang, Qian, Liu, Zhu, Yang, Han: "Therapeutic potential of NaoXinTong Capsule on the developed diabetic nephropathy in db/db mice." in: Biomedicine & pharmacotherapy, Vol. 118, pp. 109389, (2020) (PubMed).

Santos, de Bem, da Costa, de Carvalho, de Medeiros, Silva, Romão, de Andrade Soares, Ognibene, de Moura, Resende: "Açaí seed extract prevents the renin-angiotensin system activation, oxidative stress and inflammation in white adipose tissue of high-fat diet-fed mice." in: Nutrition research (New York, N.Y.), Vol. 79, pp. 35-49, (2020) (PubMed).

Ore, Ugbaja, Adeogun, Akinloye: "An albino mouse model of nonalcoholic fatty liver disease induced using high-fat liquid "Lieber-DeCarli" diet: a preliminary investigation." in: Porto biomedical journal, Vol. 5, Issue 4, pp. e071, (2020) (PubMed).

Ji, Qin, Ren, Lu, Wang, Dai, Zhou, Huang, Xu, Chen, Wu, Song, Shen, Hu, Miao, Xia, Wang: "Mitochondria-related miR-141-3p contributes to mitochondrial dysfunction in HFD-induced obesity by inhibiting PTEN." in: Scientific reports, Vol. 5, pp. 16262, (2016) (PubMed).

Target Details for Insulin

Target: Insulin (INS)

Alternative Name: Insulin (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: IDDM2, ILPR, IRDN, MODY10

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