Rat (Rattus) Insulin ELISA Kit

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Antigen
Insulin (INS) ELISA Kits
  • ins1
  • xins
  • ins1-a
  • Insulin
  • IDDM2
  • ILPR
  • IRDN
  • MODY10
  • AA986540
  • Ins-2
  • InsII
  • Mody
  • Mody4
  • proinsulin
  • zgc:109842
  • insulin
  • insulin precursor
  • insulin II
  • insulin-like
  • preproinsulin
  • ins
  • PIN
  • INS
  • Ins
  • Ins2
  • LOC100060077
Reactivity
Rat (Rattus)
Alternatives
Kits with alternative reactivity to:
67
33
31
24
17
16
14
14
10
9
8
7
6
4
3
1
1
Method Type
Competition ELISA
Detection Range
123.5-10000 pg/mL
Minimum Detection Limit
123.5 pg/mL
Application
ELISA
Options
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Catalog No. ABIN416266
$ 576.00
Plus shipping costs $45.00
Relevance Score ABIN Method Type Sample Type Detection Range Detection Minimum Supplier References Details
10.541325 ABIN1979556 Sandwich ELISA Serum, Plasma, Cell Culture Supernatant 5-300 μIU/mL 5 μIU/mL Log in to see 5
10.541325 ABIN366470 Sandwich ELISA Serum, Plasma, Cell Culture Supernatant 15.6-1000 nlU/mL 15.6 nlU/mL Log in to see 11
10.541325 ABIN2703271 Sandwich ELISA Cell Culture Supernatant, Plasma, Serum 4-300 μIU/mL 4 μIU/mL Log in to see
10.541325 ABIN2703269 Sandwich ELISA Cell Culture Supernatant, Plasma, Serum 4.5-300 μIU/mL 4.5 μIU/mL Log in to see
10.541325 ABIN366486 Sandwich ELISA Serum, Plasma, Cell Culture Supernatant, Tissue Homogenate 1.56-100 pg/mL 1.56 pg/mL Log in to see 1
10.541325 ABIN1116659 Sandwich ELISA Serum, Plasma, Biological Fluids 0.313-20 ng/mL 0.313 ng/mL Log in to see
10.541325 ABIN1873118 Competition ELISA Plasma, Cell Lysate, Cell Culture Supernatant, Serum, Tissue Homogenate, Biological Fluids 78.1-20.000 pg/mL 78.1 pg/mL Log in to see
1 ABIN1028897 Competition ELISA Serum, Plasma, Tissue Homogenate, Cell Lysate, Cell Culture Supernatant, Biological Fluids 140.62-9000 pg/mL 140.62 pg/mL Log in to see
1 ABIN5655382 Competition ELISA Biological Fluids, Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate 123.5 pg/mL - 10000 pg/mL 123.5 pg/mL Log in to see
1 ABIN5655381 Competition ELISA Biological Fluids, Cell Culture Supernatant, Cell Lysate, Plasma, Serum, Tissue Homogenate 78.12 pg/mL - 20000 pg/mL 78.12 pg/mL Log in to see
1 ABIN771959 Competition ELISA Serum, Plasma, Cell Culture Supernatant, Tissue Homogenate, Body Fluids 0.5-10 ng/mL 0.5 ng/mL Log in to see
1 ABIN5591738 Competition ELISA Biological Fluids, Plasma, Serum, Tissue Homogenate 0.625-40 μIU/mL 0.625 μIU/mL Log in to see
1 ABIN1813713 Sandwich ELISA Body Fluids, Tissue Lysate, Cell Culture Supernatant, Serum Log in to see 3
1 ABIN1057579 Sandwich ELISA Serum, Cell Culture Supernatant, Body Fluids, Tissue Homogenate, Plasma 5.0-100 pM/L 5.0 pM/L Log in to see
1 ABIN5520039 Sandwich ELISA Plasma, Serum, Biological Fluids, Tissue Homogenate 0.313-20 ng/mL 0.313 ng/mL Log in to see
1 ABIN772198 Serum, Plasma, Cell Culture Supernatant, Tissue Homogenate, Body Fluids Log in to see
1 ABIN4993536 Sandwich ELISA Plasma, Serum, Biological Fluids 3.125-200 ng/mL 3.125 ng/mL Log in to see
1 ABIN5520038 Sandwich ELISA Plasma, Serum, Biological Fluids, Tissue Homogenate 78.125-5000 pg/mL 78.125 pg/mL Log in to see
1 ABIN1774778 Serum, Plasma 0-100 μIU/mL 0 μIU/mL Log in to see 2
1 ABIN1846977 Serum, Plasma, Biological Fluids 0-140 μIU/mL 0 μIU/mL Log in to see

General

Antigen Insulin (INS) ELISA Kits
Reactivity Rat (Rattus)
Kits with alternative reactivity to:
(67), (33), (31), (24), (17), (16), (14), (14), (10), (9), (8), (7), (6), (4), (3), (1), (1)
Method Type Competition ELISA
Detection Range 123.5-10000 pg/mL
Minimum Detection Limit 123.5 pg/mL
Application ELISA
Pubmed 16 references available
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Product Details Insulin ELISA Kit

Target details Application Details Handling References for Insulin Kit (ABIN416266) Images
Purpose The kit is a competitive inhibition enzyme immunoassay technique for the in vitro quantitative measurement of INS in Serum,Plasma,Biological Fluids
Sample Type Serum, Plasma, Tissue Homogenate, Cell Lysate, Cell Culture Supernatant, Biological Fluids
Detection Method Colorimetric
Analytical Method Quantitative
Specificity

This assay has high sensitivity and excellent specificity for detection of Insulin (INS).
No significant cross-reactivity or interference between Insulin (INS) and analogues was observed.

Cross-Reactivity (Details) No significant cross-reactivity or interference between Insulin (INS) and analogues was observed.
Sensitivity 49.3 pg/mL
Components
  • Pre-coated, ready to use 96-well strip plate
  • Plate sealer for 96 wells
  • Standard Diluent
  • Assay Diluent A
  • Assay Diluent B
  • Stop Solution
  • Standard
  • Detection Reagent A
  • Detection Reagent B
  • TMB Substrate
  • Wash Buffer (30 × concentrate)
  • Instruction manual
Material not included
  • Microplate reader with 450 nm filter.
  • Precision single or multi-channel pipettes and disposable tips.
  • Eppendorf Tubes for diluting samples.
  • Deionized or distilled water.
  • Absorbent paper for blotting the microtiter plate.
  • Container for Wash Solution

Target details

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Antigen
Alternative Name INS (INS ELISA Kit Abstract)
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

Application Details

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Application Notes
  • Limited by the current condition and scientific technology, we cannot completely conduct the comprehensive identification and analysis on the raw material provided by suppliers. So there might be some qualitative and technical risks to use the kit.
  • The final experimental results will be closely related to validity of the products, operation skills of the end users and the experimental environments. Please make sure that sufficient samples are available.
  • Kits from different batches may be a little different in detection range, sensitivity and color developing time.
  • Do not mix or substitute reagents from one kit lot to another. Use only the reagents supplied by manufacturer.
  • Protect all reagents from strong light during storage and incubation. All the bottle caps of reagents should be covered tightly to prevent the evaporation and contamination of microorganism.
  • There may be some foggy substance in the wells when the plate is opened at the first time. It will not have any effect on the final assay results. Do not remove microtiter plate from the storage bag until needed.
  • Wrong operations during the reagents preparation and loading, as well as incorrect parameter setting for the plate reader may lead to incorrect results. A microplate plate reader with a bandwidth of 10nm or less and an optical density range of 0-3 O.D. or greater at 450 ± 10nm wavelength is acceptable for use in absorbance measurement. Please read the instruction carefully and adjust the instrument prior to the experiment.
  • Even the same operator might get different results in two separate experiments. In order to get better reproducible results, the operation of every step in the assay should be controlled. Furthermore, a preliminary experiment before assay for each batch is recommended.
  • Each kit has been strictly passed Q.C test. However, results from end users might be inconsistent with our in-house data due to some unexpected transportation conditions or different lab equipments. Intra-assay variance among kits from different batches might arise from above factors, too.
  • Kits from different manufacturers for the same item might produce different results, since we have not compared our products with other manufacturers.
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 This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific to Insulin (INS) has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled Insulin (INS) and unlabeled Insulin (INS) (Standards or samples) with the pre-coated antibody specific to Insulin (INS). After incubation the unbound conjugate is washed off. Next, avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The amount of bound HRP conjugate is reverse proportional to the concentration of Insulin (INS) in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of Insulin (INS) in the sample.
Reagent Preparation
  • Bring all kit components and samples to room temperature (18-25°C) before use.
  • Standard - Reconstitute the Standard with 2.0mL 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 9,000pg/mL. Please prepare 5 tubes containing 0.6mL Standard Diluent and produce a triple dilution series. Mix each tube thoroughly before the next transfer. Set up 5 points of diluted standard such as 9,000pg/mL, 3,000pg/mL, 1,000pg/mL, 333.3pg/mL, 111.1pg/mL, and the last EP tubes with Standard Diluent is the blank as 0pg/mL.
  • Assay Diluent A and Assay Diluent B - Dilute 6mL of Assay Diluent A or B Concentrate(2×) with 6mL of deionized or distilled water to prepare 12mL of Assay Diluent A or B. (In fact, more than 6mL Assay Diluent A and Assay Diluent B are contained in the bottles. Therefore, in every test, please precisely pipette required amount of Diluent and make double dilution in a new container. The prepared working dilution cannot be frozen.)
  • Detection Reagent A and Detection Reagent B - Briefly spin or centrifuge the stock Detection A and Detection B before use. Dilute to the working concentration with working Assay Diluent A(1:200) and Assay Diluent B(1:100), respectively.
  • Wash Solution - Dilute 20mL of Wash Solution concentrate (30×) with 580mL of deionized or distilled water to prepare 600mL of Wash Solution (1×).
  • TMB substrate - Aspirate the needed dosage of the solution with sterilized tips and do not dump the residual solution into the vial again.
Note:
  • Making serial dilution in the wells directly is not permitted.
  • Prepare standard within 15 minutes before assay. Please do not dissolve the reagents at 37°C directly.
  • Detection Reagent A and B are sticky solutions, therefore, slowly pipette them to reduce the volume errors.
  • 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 once pipetting.
  • The reconstituted Standards, Detection Reagent A and Detection Reagent B can be used only once.
  • If crystals have formed in the Wash Solution concentrate (30×), warm to room temperature and mix gently until the crystals are completely dissolved.
  • Contaminated water or container for reagent preparation will influence the detection result.
Sample Collection Serum: Allow samples to clot for two hours at room temperature or overnight at 4°C before centrifugation for 20 minutes at approximately 1000 × g. Assay immediately or store samples in aliquot at -20°C or -80°C. Avoid repeated freeze/thaw cycles.

Plasma: Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples for 15 minutes at 1000 × g within 30 minutes of collection. Remove plasma and assay immediately or store samples in aliquot at -20°C or -80°C. Avoid repeated freeze/thaw cycles.

Biological Fluids: Centrifuge samples for 20 minutes at 1000 × g. Remove particulates and assay immediately or store samples in aliquot at -20 °C or -80 °C for later use. Avoid repeated freeze/thaw cycles.
Sample Preparation
  • We are only responsible for the kit itself, but not for the samples consumed during the assay. The user should calculate the possible amount of the samples used in the whole test. Please reserve sufficient samples in advance.
  • Please predict the concentration before assaying. If values for these are not within the range of the standard curve, users must determine the optimal sample dilutions for their particular experiments. Sample should be diluted by 0.01mol/L PBS(PH=7.0-7.2).
  • If the samples are not indicated in the manual, a preliminary experiment to determine the validity of the kit is necessary.
  • Tissue or cell extraction samples prepared by chemical lysis buffer may cause unexpected ELISA results due to the impacts from certain chemicals.
  • Due to the possibility of mismatching between antigen from other origin and antibody used in our kits (e.g.antibody targets conformational epitope rather than linear epitope), some native or recombinant proteins from other manufacturers may not be recognized by our products.
  • Influenced by the factors including cell viability, cell number or sampling time, samples from cell culture supernatant may not be detected by the kit.
  • Fresh samples without long time storage is recommended for the test. Otherwise, protein degradation and denaturalization may occur in those samples and finally lead to wrong results.
Assay Procedure
  1. Prepare all reagents, samples and standards,
  2. Add 50μL standard or sample to each well.
    And 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.
Calculation of Results This assay employs the competitive inhibition enzyme immunoassay technique, so there is an inverse correlation between INS concentration in the sample and the assay signal intensity. Average the duplicate readings for each standard, control, and samples. Create a standard curve on log-log or semi-log graph paper, with the log of INS concentration on the y-axis and absorbance on the x-axis. Draw the best fit straight line through the standard points and it can be determined by regression analysis. Using some plot software, for instance, curve expert 1.30, is also recommended. If samples have been diluted, the concentration read from the standard curve must be multiplied by the dilution factor.
In order to make the calculation easier, we plot the O.D. value of the standard (X-axis) against the log of concentration of the standard (Y-axis), although concentration is the independent variable and O.D. value is the dependent variable. The O.D. values of the standard curve may vary according to the conditions of assay performance (e.g. operator, pipetting technique, washing technique or temperature effects). Typical standard curve below is provided for reference only.
Assay Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Insulin (INS) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Insulin (INS) 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

Handling

Product Details Insulin ELISA Kit Target details Application Details References for Insulin Kit (ABIN416266) Images back to top
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.
Handling Advice

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5 % within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Storage 4 °C
Storage Comment
  • For unopened kit: All the reagents should be kept according to the labels on vials. The Standard, Detection Reagent A, Detection Reagent B and the 96-well strip plate should be stored at -20°C upon receipt while the others should be at 4°C.
  • For opened kit: When the kit is opened, the remaining reagents still 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 reseal along entire edge of zip-seal.
    Note: It is highly recommended to use the remaining reagents within 1 month provided this is within the expiration date of the kit.
  • For ELISA kit, 1 day storage at 37°C can be considered as 2 months at 4°C, which means 3 days at 37°C equaling 6 months at 4°C.
Expiry Date 6 months

References for Insulin Kit (ABIN416266)

Product Details Insulin ELISA Kit Target details Application Details Handling Images back to top
Product cited in:

Zhang, Hu, Meng, Sun, Xu, Zhang, Cui, Morina, Li, Li, Wu, Brännström, Shao, Billig: "Metformin Ameliorates Uterine Defects in a Rat Model of Polycystic Ovary Syndrome." in: EBioMedicine, Vol. 18, pp. 157-170, 2017

Wang, Zhou, Quach, Lu, Gao, Xu, Zhu: "Effect of Sleeve Gastrectomy Plus Side-to-Side Jejunoileal Anastomosis for Type 2 Diabetes Control in an Obese Rat Model." in: Obesity surgery, Vol. 26, Issue 4, pp. 797-804, 2016

Zhou, Guo, Huang, Zhu, Fan, Wang, Wang, Zhu, Xu, Wu, Lu, Wang: "The dynamic three-dimensional culture of islet-like clusters in decellularized liver scaffolds." in: Cell and tissue research, Vol. 365, Issue 1, pp. 157-71, 2016

Olatunji, Omolekulo, Usman, Kim: "Improvement of oral contraceptive-induced glucose dysregulation and dyslipidemia by valproic acid is independent of circulating corticosterone." in: Archives of physiology and biochemistry, Vol. 122, Issue 3, pp. 123-9, 2016

Fang, Yu, He, Guo, Huang, Kong, Shi, Zhu, Bo, Zhang: "Central injection of GALR1 agonist M617 attenuates diabetic rat skeletal muscle insulin resistance through the Akt/AS160/GLUT4 pathway." in: Mechanisms of ageing and development, 2016

Ali, El-Sayyad, Moftah, Chilibeck: "Structural and functional abnormalities of hepatic tissues in male Wistar rats fed hyperwhey and super amino anabolic protein." in: Nutrition (Burbank, Los Angeles County, Calif.), Vol. 32, Issue 7-8, pp. 840-8, 2016

Lavet, Martin, Linossier, Vanden Bossche, Laroche, Thomas, Gerbaix, Ammann, Fraissenon, Lafage-Proust, Courteix, Vico: "Fat and Sucrose Intake Induces Obesity-Related Bone Metabolism Disturbances: Kinetic and Reversibility Studies in Growing and Adult Rats." in: Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research, Vol. 31, Issue 1, pp. 98-115, 2016

Olatunji, Usman, Seok, Kim: "Activation of cardiac renin-angiotensin system and plasminogen activator inhibitor-1 gene expressions in oral contraceptive-induced cardiometabolic disorder." in: Archives of physiology and biochemistry, pp. 1-8, 2016

Zhang, Sun, Sun, Meng, Hu, Li, Li, Wu, Brännström, Shao, Billig: "Molecular characterization of insulin resistance and glycolytic metabolism in the rat uterus." in: Scientific reports, Vol. 6, pp. 30679, 2016

Ding, Zhang, Dong, Ding, Huang, Zhu, Hutchinson, Gao, Zhang: "Adiponectin protects the rats liver against chronic intermittent hypoxia induced injury through AMP-activated protein kinase pathway." in: Scientific reports, Vol. 6, pp. 34151, 2016

Zhang, Li, Li, Zhang, Chen, Liu, Zhang, Zhang, Yang, Hu, Wu, Li, Ju, Yang: "The anti-hyperglycemic efficacy of a lipid-lowering drug Daming capsule and the underlying signaling mechanisms in a rat model of diabetes mellitus." in: Scientific reports, Vol. 6, pp. 34284, 2016

Abu, Samat, Kamarapani, Nor Hussein, Wan Ismail, Hassan: "Tinospora crispa Ameliorates Insulin Resistance Induced by High Fat Diet in Wistar Rats." in: Evidence-based complementary and alternative medicine : eCAM, Vol. 2015, pp. 985042, 2015

Nascimento da Silva, Azevedo de Jesuz, De Salvo Castro, Soares da Costa, Teles Boaventura, Blondet de Azeredo: "Effect of the “protein diet” and bone tissue." in: Nutrición hospitalaria, Vol. 29, Issue 1, pp. 140-5, 2014

Yang, Luo, Zhou, Lv, Liu, Zhang, Gao, Chen, Xia, Luo, Cheng, Li: "Rosiglitazone inhibits expression and secretion of PEDF in adipose tissue and liver of male SD rats via a PPAR-? independent mechanism." in: Endocrinology, Vol. 155, Issue 3, pp. 941-50, 2014

Mohammadi, Gholamhoseinian, Fallah: "Zataria multiflora increases insulin sensitivity and PPAR? gene expression in high fructose fed insulin resistant rats." in: Iranian journal of basic medical sciences, Vol. 17, Issue 4, pp. 263-70, 2014

Tinkov, Popova, Polyakova, Nikonorov: "Perinatal low-dose iron treatment influences susceptibility to diet-induced adipogenesis in early-aged male Wistar rats." in: Biometals : an international journal on the role of metal ions in biology, biochemistry, and medicine, Vol. 27, Issue 2, pp. 293-303, 2014

Images

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Supplier Images
ELISA image for Insulin (INS) ELISA Kit (ABIN416266) Insulin (INS) ELISA Kit