Insulin ELISA Kit

Catalog No. ABIN416266

Product Details Insulin ELISA Kit

Target: Insulin (INS)

Reactivity: Rat

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 rat 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).
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, 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

Application Details

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:

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.

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

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 ELISA Kit (ABIN416266)

Yan, Chen, Zhu, Lin, Sun, Liu, Pan, Wang, Yang, Liu, Gong: "The Changes of Serum Metabolites in Diabetic GK Rats after Ileal Transposition Surgery." in: Obesity surgery, Vol. 29, Issue 3, pp. 882-890, (2020) (PubMed).

Dolo, Shao, Li, Zhu, Yao, Wang: "The Effect of Gastric Bypass with a Distal Gastric Pouch on Glucose Tolerance and Diabetes Remission in Type 2 Diabetes Sprague-Dawley Rat Model." in: Obesity surgery, Vol. 29, Issue 6, pp. 1889-1900, (2020) (PubMed).

Wang, Ai, Hai, Cao, Li, Li, Li: "Tempol alleviates chronic intermittent hypoxia-induced pancreatic injury through repressing inflammation and apoptosis." in: Physiological research, Vol. 68, Issue 3, pp. 445-455, (2020) (PubMed).

Zhai, Li, Zhang, Ma, Ma, Zhang, Zou, Li, Ma, Li: "Coptisine ameliorates renal injury in diabetic rats through the activation of Nrf2 signaling pathway." in: Naunyn-Schmiedeberg's archives of pharmacology, Vol. 393, Issue 1, pp. 57-65, (2020) (PubMed).

Li, Yan, Jing, Pan, Wang, Jiang, Li, Wang, Dong, Wang, Zhang, Meng, Li, Liu, Gao, Qu: "Duodenal-Jejunal Bypass Ameliorates Type 2 Diabetes Mellitus by Activating Insulin Signaling and Improving Glucose Utilization in the Brain." in: Obesity surgery, Vol. 30, Issue 1, pp. 279-289, (2020) (PubMed).

Dolo, Huang, Widjaja, Li, Zhu, Yao, Hong: "Distal gastric mucosa ablation induces significant weight loss and improved glycemic control in type 2 diabetes Sprague-Dawley rat model." in: Surgical endoscopy, Vol. 34, Issue 10, pp. 4336-4346, (2020) (PubMed).

Ren, Shao, Wu, Ma, Lv, Wang: "Metformin alleviates oxidative stress and enhances autophagy in diabetic kidney disease via AMPK/SIRT1-FoxO1 pathway." in: Molecular and cellular endocrinology, Vol. 500, pp. 110628, (2020) (PubMed).

Mu, Xin, Zhang, Wang, Ning, Meng: "Beneficial effects of Aronia melanocarpa berry extract on hepatic insulin resistance in type 2 diabetes mellitus rats." in: Journal of food science, Vol. 85, Issue 4, pp. 1307-1318, (2020) (PubMed).

Huang, Huang, Li, Bai, Hao, Wu, Yi, Xu, Pan, Zhang: "Electroacupuncture improves cognitive deficits and insulin resistance in an OLETF rat model of Al/D-gal induced aging model via the PI3K/Akt signaling pathway." in: Brain research, Vol. 1740, pp. 146834, (2020) (PubMed).

Youssef, Morsy, El-Shahat, Shams, Abd-Elhady: "The neuroprotective effect of simvastatin on the cerebellum of experimentally-induced diabetic rats through klotho upregulation: An immunohistochemical study." in: Journal of chemical neuroanatomy, Vol. 108, pp. 101803, (2020) (PubMed).

Guo, He, Fang, Bo, Zhang: "Activation of central galanin receptor 2 mitigated insulin resistance in adipocytes of diabetic rats." in: Journal of endocrinological investigation, (2020) (PubMed).

Samaha, Said, Salem: "Modulatory role of imatinib mesylate on pancreatic β-cells' secretory functions in an STZ rat model of diabetes mellitus." in: Chemico-biological interactions, Vol. 328, pp. 109197, (2020) (PubMed).

Ahmed, Salem, Seif El-Din, El-Lakkany, Ahmed, Nasr, Hammam, Botros, Saleh: "Gut microbiota modulation as a promising therapy with metformin in rats with non-alcoholic steatohepatitis: Role of LPS/TLR4 and autophagy pathways." in: European journal of pharmacology, Vol. 887, pp. 173461, (2020) (PubMed).

Tian, Zhang, Li, Li, Wang, Zhang, Ma: "Palmatine ameliorates high fat diet induced impaired glucose tolerance." in: Biological research, Vol. 53, Issue 1, pp. 39, (2020) (PubMed).

Lange, Osinsky: "Aiming at ecological validity-Midfrontal theta oscillations in a toy gun shooting task." in: The European journal of neuroscience, (2020) (PubMed).

Dong, Hou, Liu, Tao, Zhang, Zhao, Song: "Regulation of insulin resistance by targeting the insulin-like growth factor 1 receptor with microRNA-122-5p in hepatic cells." in: Cell biology international, Vol. 43, Issue 5, pp. 553-564, (2019) (PubMed).

Yusoff, Lim, Al-Hindi, Abdul Razak, Widyawati, Anggraini, Ahmad, Asmawi: "Nypa fruticans Wurmb. Vinegar's Aqueous Extract Stimulates Insulin Secretion and Exerts Hepatoprotective Effect on STZ-Induced Diabetic Rats." in: Nutrients, Vol. 9, Issue 9, (2018) (PubMed).

Stygar, Sawczyn, Skrzep-Poloczek, Owczarek, Matysiak, Michalski, Mielańczyk, Bażanów, Ziora, Choręza, Doleżych, Karcz: "The Effects of Duodenojejunal Omega Switch in Combination with High-Fat Diet and Control Diet on Incretins, Body Weight, and Glucose Tolerance in Sprague-Dawley Rats." in: Obesity surgery, Vol. 28, Issue 3, pp. 748-759, (2018) (PubMed).

Kołodziej, Maciejczyk, Niklińska, Waszkiel, Żendzian-Piotrowska, Żukowski, Zalewska: "Chronic high-protein diet induces oxidative stress and alters the salivary gland function in rats." in: Archives of oral biology, Vol. 84, pp. 6-12, (2018) (PubMed).

Zhao, Zhou, Chen, Liu, Chu, Zhang: "Beneficial effects of Heqi san on rat model of polycystic ovary syndrome through the PI3K/AKT pathway." in: Daru : journal of Faculty of Pharmacy, Tehran University of Medical Sciences, Vol. 25, Issue 1, pp. 21, (2018) (PubMed).

Target Details for Insulin

Target: Insulin (INS)

Alternative Name: 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

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