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Rat INS(Insulin) ELISA Kit

Citations(25)Uniprot : P01322,P01323

  • Cat.No.:E-EL-R2466

  • Reactivity: Rat
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  • 96T
  • 48T
  • 24T
  • 96T*5
  • 96T*10
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Product Details

Properties

Assay type Sandwich-ELISA
Format 96T
Assay time 3.5h
Detection range 0.31-20 ng/mL
Sensitivity 0.19 ng/mL
Sample type &Sample volume cell culture supernatant and tissue homogenate; 100μL
Specificity This kit recognizes Rat INS in samples. No significant cross-reactivity or interference between Rat INS and analogues was observed.
Reproducibility Both intra-CV and inter-CV are < 10%.
Application This ELISA kit applies to the in vitro quantitative determination of Rat INS concentrations in cell culture supernatant and tissue homogenate.

Test Principle

This ELISA kit uses the Sandwich-ELISA principle. The micro ELISA plate provided in this kit has been pre-coated with an antibody specific to Rat INS. Standards or samples are added to the micro ELISA plate wells and combined with the specific antibody. Then a biotinylated detection antibody specific for Rat INS and Avidin-Horseradish Peroxidase (HRP) conjugate are added successively to each micro plate well and incubated. Free components are washed away. The substrate solution is added to each well. Only those wells that contain Rat INS, biotinylated detection antibody and Avidin-HRP conjugate will appear blue in color. The enzyme-substrate reaction is terminated by the addition of stop solution and the color turns yellow. The optical density (OD) is measured spectrophotometrically at a wavelength of 450 nm ± 2 nm. The OD value is proportional to the concentration of Rat INS. You can calculate the concentration of Rat INS in the samples by comparing the OD of the samples to the standard curve.

Kit components & Storage

An unopened kit can be stored at 2-8℃ for 1 month. If the kit is not supposed to be used within 1 month, store the components separately according to the following conditions once the kit is received.
ItemSpecificationsStorage
Micro ELISA Plate(Dismountable) 96T: 8 wells ×12 strips
48T: 8 wells ×6 strips 		-20℃, 6 months
Reference Standard 96T: 2 vials
48T: 1 vial
Concentrated Biotinylated Detection Ab (100×) 96T: 1 vial, 120 μL
48T: 1 vial, 60 μL
Concentrated HRP Conjugate (100×) 96T: 1 vial, 120 μL
48T: 1 vial, 60 μL 		-20℃(Protect from light), 6 months
Reference Standard & Sample Diluent 1 vial, 20 mL 2-8°C, 6 months
Biotinylated Detection Ab Diluent 1 vial, 14 mL
HRP Conjugate Diluent 1 vial, 14 mL
Concentrated Wash Buffer (25×) 1 vial, 30 mL
Substrate Reagent 1 vial, 10 mL 2-8℃(Protect from light)
Stop Solution 1 vial, 10 mL 2-8°C
Plate Sealer 5 pieces
Manual 1 copy
Certificate of Analysis 1 copy

Technical Data

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, mid range and high level Rat INS were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, mid range and high level Rat INS were tested on 3 different plates, 20 replicates in each plate.

  Intra-assay Precision Inter-assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (ng/mL) 0.99 2.68 6.73 1.00 2.82 6.90
Standard deviation 0.06 0.14 0.25 0.07 0.16 0.35
CV (%) 6.06 5.22 3.71 7.00 5.67 5.07

Recovery

The recovery of Rat INS spiked at three different levels in samples throughout the range of the assay was evaluated in various matrices.

Sample Type Range (%) Average Recovery (%)
Cell culture media (n=8) 86-98 92

Linearity

Samples were spiked with high concentrations of Rat INS and diluted with Reference Standard & Sample Diluent to produce samples with values within the range of the assay.

Target Information

Database LinksSwissProt:   P01322  P01323
SynonymsIDDM2, ILPR, IRDN, MODY10
Research AreaCancer

Assay Procedures

elisa assay procedure 1

1. Add 100μL standard or sample to the wells. Incubate for 90 min at 37°C
elisa assay procedure 2

2. Discard the liquid, immediately add 100μL Biotinylated Detection Ab working solution to each well. Incubate for 60 min at 37°C
elisa assay procedure 3

3. Aspirate and wash the plate for 3 times
elisa assay procedure 4

4. Add 100μL HRP conjugate working solution. Incubate for 30 min at 37°C. Aspirate and wash the plate for 5 times
elisa assay procedure 5

5. Add 90μL Substrate Reagent. Incubate for 15 min at 37°C
elisa assay procedure 6

6. Add 50μL Stop Solution
elisa assay procedure 7

7. Read the plate at 450nm immediately. Calculation of the results

Citations

  1. FOOD RESEARCH INTERNATIONAL (2021) IF: 6.475
    Potential mechanistic pathways underlying intestinal and hepatic effects of kefir in high-fructose-fed rats

    DOI: 10.1016/j.foodres.2021.110287

    PMID: 33992387

    		 Sample: Tissue homogenate
  2. Frontiers in Cell and Developmental Biology (2021) IF: 6.684
    GLP-1-Induced AMPK Activation Inhibits PARP-1 and Promotes LXR-Mediated ABCA1 Expression to Protect Pancreatic β-Cells Against Cholesterol-Induced Toxicity Through Cholesterol Efflux

    DOI: 10.3389/fcell.2021.646113
    Sample: INS-1 cells
  3. Viruses-Basel (2022) IF: 5.818
    Replication Activities of Major 5′ Terminally Deleted Group-B Coxsackievirus RNA Forms Decrease PCSK2 mRNA Expression Impairing Insulin Maturation in Pancreatic Beta Cells

    DOI: 10.3390/v14122781
    Sample: INS-1 cells
  4. BRAIN RESEARCH BULLETIN (2021) IF: 4.079
    Role of protein tyrosine phosphatase 1B inhibitor in central insulin resistance and associated cognitive deficits

    DOI: 10.1016/j.brainresbull.2021.02.026

    PMID: 33684458

    		 Sample: Tissue homogenate
  5. MOLECULAR AND CELLULAR ENDOCRINOLOGY (2021) IF: 4.102
    Heme Oxygenase-1 (HMOX-1) and inhibitor of differentiation proteins (ID1, ID3) are key response mechanisms against iron-overload in pancreatic β-cells

    DOI: 10.1016/j.mce.2021.111462

    PMID: 34547407

    		 Sample: Pancreatic β cell
  6. EXPERIMENTAL CELL RESEARCH (2022) IF: 4.145
    Sox9 is required in regeneration of pancreatic β cells following injury

    DOI: 10.1016/j.yexcr.2022.113406
    Sample: RIN-m5F cell
  7. AMERICAN JOURNAL OF PHYSIOLOGY-ENDOCRINOLOGY AND METABOLISM (2021) IF: 4.31
    Bee bread mitigates downregulation of steroidogenic genes, decreased spermatogenesis, and epididymal oxidative stress in male rats fed with high-fat diet

    DOI: 10.1152/ajpendo.00093.2021

    PMID: 34229480

    		 Sample: serum,testis
  8. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES (2020) IF: 4.556
    MKP-5 Relieves Lipotoxicity-Induced Islet β-Cell Dysfunction and Apoptosis via Regulation of Autophagy

    DOI: 10.3390/ijms21197161

    PMID: 32998359

    		 Sample: Cell lysate
  9. Biomolecules (2018) IF: 4.879
    Methylation and Acetylation Enhanced the Antidiabetic Activity of Some Selected Flavonoids: In Vitro, Molecular Modelling and Structure Activity Relationship-Based Study

    DOI: 10.3390/biom8040149

    PMID: 30445784

    		 Sample: Cell culture medium
  10. Chinese Journal of Natural Medicines (2014) IF: 3
    Modified Si-Miao-San ameliorates pancreatic B cell dysfunction by inhibition of reactive oxygen species-associated inflammation through AMP-kinase activation

    DOI: 10.1016/S1875-5364(14)60043-1
    Sample: Cell culture supernatant
  11. KIDNEY & BLOOD PRESSURE RESEARCH (2017) IF: 3.104
    The Ameliorative Effect of Pectin-Insulin Patch On Renal Injury in Streptozotocin-Induced Diabetic Rats

    DOI: 10.1159/000480395
    Sample: Urine,Tissue homogenate,Plasma
  12. PFLUGERS ARCHIV-EUROPEAN JOURNAL OF PHYSIOLOGY (2020) IF: 3.158
    Nitric oxide interacts with cholinoceptors to modulate insulin secretion by pancreatic β cells

    DOI: 10.1007/s00424-020-02443-9

    PMID: 32803305

    		 Sample: Cell culture supernatant
  13. CELL TRANSPLANTATION (2020) IF: 3.341
    Bone Marrow-Derived Mesenchymal Stem Cells Restored High-Fat-Fed Induced Hyperinsulinemia in Rats at Early Stage of Type 2 Diabetes Mellitus:

    DOI: 10.1177/0963689720904628
    Sample: Serum,plasma
  14. LIFE SCIENCES (2019) IF: 3.448
    Effects of noopept on cognitive functions and pubertal process in rats with diabetes

    DOI: 10.1016/j.lfs.2019.116698

    PMID: 31356906

    		 Sample: Serum,Tissue homogenate
  15. MOLECULAR AND CELLULAR ENDOCRINOLOGY (2019) IF: 3.693
    Orphan G-protein coupled receptor 183 (GPR183) potentiates insulin secretion and prevents glucotoxicity-induced β-cell dysfunction

    DOI: 10.1016/j.mce.2019.110592
    Sample: INS-1 (832/13) cells
  16. BIOSCIENCE REPORTS (2018) IF: 3.84
    The Chinese medicine Chai Hu Li Zhong Tang protects against non-alcoholic fatty liver disease by activating AMPKα

    DOI: 10.1042/BSR20180644
    Sample: Cell culture supernatant
  17. EXPERIMENTAL AND CLINICAL ENDOCRINOLOGY & DIABETES (2020) IF: 2.058
    Let7b-5p is Upregulated in the Serum of Emirati Patients with Type 2 Diabetes and Regulates Insulin Secretion in INS-1 Cells

    DOI: 10.1055/a-1261-5282
    Sample: INS-1 (832/13) cell
  18. Veterinary Sciences (2021) IF: 2.304
    Protective and Therapeutic Effects of Orlistat on Metabolic Syndrome and Oxidative Stress in High-Fat Diet-Induced Metabolic Dysfunction-Associated Fatty Liver Disease (MAFLD) in Rats: Role on Nrf2 Activation

    DOI: 10.3390/vetsci8110274
  19. Biomed Research International (2017) IF: 2.476
    Cotransplantation of Mesenchymal Stem Cells and Immature Dendritic Cells Potentiates the Blood Glucose Control of Islet Allografts

    DOI: 10.1155/2017/4107943
    Sample: Cell culture supernatant
  20. GENE (2019) IF: 2.638
    GNAS gene is an important regulator of insulin secretory capacity in pancreatic β-cells

    DOI: 10.1016/j.gene.2019.144028
    Sample: INS -1 (832/13) cells
  21. Chinese Medicine (2020) IF: 2.96
    Protective effects of essential oil from Fructus Alpiniae zerumbet on retinal Müller gliosis via the PPAR-γ-p-CREB signaling pathway

    DOI: 10.1186/s13020-019-0283-4

    PMID: 31938037

    		 Sample: Serum,Cell culture medium
  22. Food Science & Nutrition (2019) IF: 1.747
    Aqueous leaf extract of Clinacanthus nutans improved metabolic indices and sorbitol‐related complications in type II diabetic rats (T2D)

    DOI: 10.1002/fsn3.988
    Sample: Tissue homogenate,Serum
  23. Experimental and Therapeutic Medicine (2020) IF: 1.785
    CGA ameliorates cognitive decline by regulating the PI3K/AKT signaling pathway and neurotransmitter systems in rats with multi-infarct dementia

    DOI: 10.3892/etm.2020.9198

    PMID: 32963600

    		 Sample: Tissue homogenate
  24. Islets (2019) IF: 1.879
    RORB and RORC associate with human islet dysfunction and inhibit insulin secretion in INS-1 cells

    DOI: 10.1080/19382014.2019.1566684
    Sample: INS-1 (832/13) cells
  25. EXPERIMENTAL AND CLINICAL ENDOCRINOLOGY & DIABETES (2019) IF: 1.927
    Reduced Expression of Chl1 gene Impairs Insulin Secretion by Down-Regulating the Expression of Key Molecules of β-cell Function

    DOI: 10.1055/a-1014-2544

    PMID: 31614370

    		 Sample: Cell culture supernatant
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