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Insulin ELISA Kit

INS Reactivity: Mouse Colorimetric Competition ELISA 123.5 pg/mL - 10000 pg/mL Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate
Catalog No. ABIN6574080
  • Target See all Insulin (INS) ELISA Kits
    Insulin (INS)
    Reactivity
    • 9
    • 8
    • 8
    • 7
    • 6
    • 4
    • 3
    • 3
    • 3
    • 3
    • 2
    • 2
    • 2
    • 1
    • 1
    Mouse
    Detection Method
    Colorimetric
    Method Type
    Competition ELISA
    Detection Range
    123.5 pg/mL - 10000 pg/mL
    Minimum Detection Limit
    123.5 pg/mL
    Application
    ELISA
    Purpose
    The kit is a competitive inhibition enzyme immunoassay technique for the in vitro quantitative measurement of insulin in mouse serum, plasma, tissue homogenates, cell lysates, cell culture supernates.

    We offer validation data (WB) for the kit components. So you can be sure to order a reliable ELISA kit product composed of high quality reagents.
    Sample Type
    Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate
    Analytical Method
    Quantitative
    Specificity
    This assay has high sensitivity and excellent specificity for detection of Insulin (INS)
    Cross-Reactivity (Details)
    No significant cross-reactivity or interference between Insulin (INS) and analogues was observed.
    Sensitivity
    48.5 pg/mL
    Components
    • Pre-coated, ready to use 96-well strip plate, flat buttom
    • Plate sealer for 96 wells
    • Reference Standard
    • Standard Diluent
    • Detection Reagent A
    • Detection Reagent B
    • Assay Diluent A
    • Assay Diluent B
    • Reagent Diluent (if Detection Reagent is lyophilized)
    • TMB Substrate
    • Stop Solution
    • Wash Buffer (30 x concentrate)
    • Instruction manual
    Top Product
    Discover our top product INS ELISA Kit
  • Comment

    Information on standard material:
    The standard might be recombinant protein or natural protein, that will depend on the specific kit. Moreover, the expression system is E.coli or yeast or mammal cell. There is 0.05% proclin 300 in the standard as preservative.

    Information on reagents:
    The stop solution used in the kit is sulfuric acid with concentration of 1 mol/L. And the wash solution is TBS. The standard diluent contains 0.02 % sodium azide, assay diluent A and assay diluent B contain 0.01% sodium azide. Some kits can contain is BSA in them.

    Information on antibodies:
    The provided antibodies and their host vary in different kits.

    Sample Volume
    50 μL
    Assay Time
    2 h
    Plate
    Pre-coated
    Protocol
    1. Prepare all reagents, samples and standards,
    2. Add 50μL standard or sample to each well.
      Then add 50μL prepared Detection Reagent A immediately.
      Shake and mix. Incubate 1 hour at 37 °C,
    3. Aspirate and wash 3 times,
    4. Add 100μL prepared Detection Reagent B. Incubate 30 minutes at 37 °C,
    5. Aspirate and wash 5 times,
    6. Add 90μL Substrate Solution. Incubate 10-20 minutes at 37 °C,
    7. Add 50μL Stop Solution. Read at 450 nm immediately.
    Reagent Preparation
    1. Bring all kit components and samples to room temperature (18-25 °C) before use. If the kit will not be used up in one time, please only take out strips and reagents for present experiment, and leave the remaining strips and reagents in required condition.
    2. Standard - Reconstitute the Standard with 0.5 mL of Standard Diluent, kept for 10 minutes at room temperature, shake gently(not to foam). The concentration of the standard in the stock solution is 10,000pg/mL. Please prepare 5 tubes containing 0.6 mL Standard Diluent and produce a triple dilution series according to the picture shown below. Mix each tube thoroughly before the next transfer. Set up 5 points of diluted standard such as 10,000pg/mL, 3,333.3pg/mL, 1,111.1pg/mL, 370.4pg/mL, 123.5pg/mL, and the last EP tubes with Standard Diluent is the blank as 0pg/mL.
    3. Detection Reagent A and Detection Reagent B - If lyophilized reconstitute the Detection Reagent A with 150μL of Reagent Diluent, kept for 10 minutes at room temperature, shake gently (not to foam). Briefly spin or centrifuge the stock Detection A and Detection B before use. Dilute them to the working concentration 100-fold with Assay Diluent A and B, respectively.
    4. Wash Solution - Dilute 20 mL of Wash Solution concentrate (30x) with 580 mL of deionized or distilled water to prepare 600 mL of Wash Solution (1x).
    5. TMB substrate - Aspirate the needed dosage of the solution with sterilized tips and do not dump the residual solution into the vial again.

    Note:

    1. Making serial dilution in the wells directly is not permitted.
    2. Prepare standard within 15 minutes before assay. Please do not dissolve the reagents at 37 °C directly.
    3. Detection Reagent A and B are sticky solutions, therefore, slowly pipette them to reduce the volume errors.
    4. Please carefully reconstitute Standards or working Detection Reagent A and B according to the instruction, and avoid foaming and mix gently until the crystals are completely dissolved. To minimize imprecision caused by pipetting, use small volumes and ensure that pipettors are calibrated. It is recommended to suck more than 10μL for one pipetting.
    5. The reconstituted Standards, Detection Reagent A and Detection Reagent B can be used only once.
    6. If crystals have formed in the Wash Solution concentrate (30x), warm to room temperature and mix gently until the crystals are completely dissolved.
    7. 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).
    Assay Precision
    Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level of target were tested 20 times on one plate, respectively.
    Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level of target were tested on 3 different plates, 8 replicates in each plate.
    CV(%) = SD/meanX100
    Intra-Assay: CV < 10%
    Inter-Assay: CV < 12%
    Restrictions
    For Research Use only
  • Precaution of Use
    The Stop Solution suggested for use with this kit is an acid solution. Wear eye, hand, face, and clothing protection when using this material.
    Storage
    4 °C/-20 °C
    Storage Comment
    1. For unopened kit: All reagents should be stored according to the labels on the vials. The Standard, Detection Reagent A, Detection Reagent B, and 96-well Strip Plate should be stored at -20 °C upon receipt, while the other reagents should be stored at 4 °C.
    2. For opened kits: the remaining reagents must be stored according to the above storage conditions. In addition, please return the unused wells to the foil pouch containing the desiccant and seal the foil pouch with the zipper.
    .
    Expiry Date
    6 months
  • Niu, Yang, Yang: "Synthesis and anti-diabetic activity of new N,N-dimethylphenylenediamine-derivatized nitrilotriacetic acid vanadyl complexes." in: Journal of inorganic biochemistry, Vol. 177, pp. 291-299, (2018) (PubMed).

    Chen, Yang, Nie, Song, Gu: "Effects of Bariatric Surgery on Change of Brown Adipocyte Tissue and Energy Metabolism in Obese Mice." in: Obesity surgery, Vol. 28, Issue 3, pp. 820-830, (2018) (PubMed).

    Tang, Tang, Wang, Ge, Zhu, Bi: "Insulin restores UCP3 activity and decreases energy surfeit to alleviate lipotoxicity in skeletal muscle." in: International journal of molecular medicine, Vol. 40, Issue 6, pp. 2000-2010, (2018) (PubMed).

    Fang, Zhang, Yu, Sheng, Shi, Zhu, Zhang, Bo: "Activiated galanin receptor 2 attenuates insulin resistance in skeletal muscle of obese mice." in: Peptides, Vol. 99, pp. 92-98, (2018) (PubMed).

    Han, Zhang, Shao, Mu, Jiao, Hou, Sun: "C1q/TNF-related protein 9 improves the anti-contractile effects of perivascular adipose tissue via the AMPK-eNOS pathway in diet-induced obese mice." in: Clinical and experimental pharmacology & physiology, Vol. 45, Issue 1, pp. 50-57, (2018) (PubMed).

    Fang, Yu, Min, Han, Shi, Zhang, Bo: "Beneficial effect of baicalin on insulin sensitivity in adipocytes of diet-induced obese mice." in: Diabetes research and clinical practice, Vol. 139, pp. 262-271, (2018) (PubMed).

    Xiao, Lou, Lu, Yang, Liu, Liu, Qi, Li: "Ginsenoside Rg5 attenuates hepatic glucagon response via suppression of succinate-associated HIF-1α induction in HFD-fed mice." in: Diabetologia, Vol. 60, Issue 6, pp. 1084-1093, (2017) (PubMed).

    Qi, Chen, Wang, Liu, Liu, Ke, Zheng, Lin, Zhou, Wu, Liu: "Subcutaneous liraglutide ameliorates methylglyoxal-induced Alzheimer-like tau pathology and cognitive impairment by modulating tau hyperphosphorylation and glycogen synthase kinase-3β." in: American journal of translational research, Vol. 9, Issue 2, pp. 247-260, (2017) (PubMed).

    Fang, Yu, Zhang, Wan, Shi, Zhu, Bo, Zhang: "Baicalin against obesity and insulin resistance through activation of AKT/AS160/GLUT4 pathway." in: Molecular and cellular endocrinology, Vol. 448, pp. 77-86, (2017) (PubMed).

    Hatem-Vaquero, Griera, García-Jerez, Luengo, Álvarez, Rubio, Calleros, Rodríguez-Puyol, Rodríguez-Puyol, De Frutos: "Peripheral insulin resistance in ILK-depleted mice by reduction of GLUT4 expression." in: The Journal of endocrinology, Vol. 234, Issue 2, pp. 115-128, (2017) (PubMed).

    Hou, Du, Han, Zhang, Jiao, Sun: "Irisin Regulates Heme Oxygenase-1/Adiponectin Axis in Perivascular Adipose Tissue and Improves Endothelial Dysfunction in Diet-Induced Obese Mice." in: Cellular physiology and biochemistry : international journal of experimental cellular physiology, biochemistry, and pharmacology, Vol. 42, Issue 2, pp. 603-614, (2017) (PubMed).

    Qi, Ke, Liu, Liao, Ke, Liu, Wang, Lin, Zhou, Wu, Chen, Liu et al.: "Subcutaneous administration of liraglutide ameliorates learning and memory impairment by modulating tau hyperphosphorylation via the glycogen synthase kinase-3β pathway in an amyloid β protein ..." in: European journal of pharmacology, Vol. 783, pp. 23-32, (2016) (PubMed).

    Hou, Liu, Han, Wang, Hou, Hou, Sun: "Irisin improves perivascular adipose tissue dysfunction via regulation of the heme oxygenase-1/adiponectin axis in diet-induced obese mice." in: Journal of molecular and cellular cardiology, Vol. 99, pp. 188-196, (2016) (PubMed).

    Guo, Yu, Li, Liu, Feng, Wang, Meng, Li, Ju, Wang: "Impaired neural stem/progenitor cell proliferation in streptozotocin-induced and spontaneous diabetic mice." in: Neuroscience research, Vol. 68, Issue 4, pp. 329-36, (2010) (PubMed).

  • Target See all Insulin (INS) ELISA Kits
    Insulin (INS)
    Abstract
    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
    UniProt
    P01326
    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
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