Insulin ELISA Kit

Catalog No. ABIN4993536

Product Details Insulin ELISA Kit

Target: Insulin (INS)

Reactivity: Rat

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 6.25 pg/mL - 400 pg/mL

Minimum Detection Limit: 6.25 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: Plasma, Serum

Analytical Method: Quantitative

Specificity: This kit recognizes natural and recombinantRatINS. No significant cross-reactivity or interference between RatINS and analogues was observed. Note: Limited by existing techniques, cross reaction may still exist, as it is impossible for us to complete the cross-reactivity detection between RatINS and all the analogues.

Sensitivity: 3.75 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

Comment:

Information on standard material:
The formulation of the standard is 0.01 M PBS. The standard contains additives (1 % BSA).

Information on reagents:
Reagents include 1 M SO₂. Azide, thimerosal, 2-mercaptoethanol (2-ME) or any other poisonous materials are not used.

Information on antibodies:
The provided antibodies and their host vary in different kits. All antibodies are affinity purified

The sensitivity of this assay, or Lower Limit of Detection (LLD) was defined as the lowest protein concentration that could be differentiated from zero. It was determined by adding two standard deviations to the mean optical density value of twenty zero standard replicates and calculating the corresponding concentration.

Sample Volume: 100 µL

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. If the kit will not be used up in one assay, please only take out the necessary strips and reagents for present experiment, and store the remaining strips and reagents at required condition.
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 400pg/mL(or add 1.0 mL of Reference Standard &Sample Diluent, let it stand for 1-2 min and then mix it thoroughly with a vortex meter of low speed. Bubbles generated during vortex could be removed by centrifuging at a relatively low speed). Then make serial dilutions as needed. The recommended dilution gradient is as follows: 400, 200, 100, 50, 25, 12.5, 6.25, 0pg/mL. Dilution method: Take 7 EP tubes, add 500μL of Reference Standard & Sample Diluent to each tube. Pipette 500μL of the 400pg/mL working solution to the first tube and mix up to produce a 200pg/mL working solution. Pipette 500μL of the solution from the former tube into the latter one according to this step. 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 Concentrated Biotinylated Detection Antibody at 800xg for 1 min, then dilute the 100x Concentrated Biotinylated Detection Antibody to 1x working solution with Biotinylated Detection Antibody Diluent(Concentrated Biotinylated Detection Antibody: Biotinylated Detection Antibody Diluent= 1: 99).
5. HRP Conjugate working solution: Calculate the required amount before the experiment (100 μL/well). In preparation, slightly more than calculated should be prepared. Centrifuge the Concentrated HRP Conjugate at 800xg for 1 min, then dilute -6- the 100x Concentrated HRP Conjugate to 1x working solution with HRP Conjugate Diluent(Concentrated HRP Conjugate: HRP Conjugate Diluent= 1: 99). 400 200 100 50 25 12.5 6.25 0

Restrictions: For Research Use only

Handling

Handling Advice: All the reagents in the kit should be stored according to the labels on vials. Unused wells should be returned to the foil pouch with the desiccant pack and resealed along entire edge of zip-seal. Substrate Reagent shouldn't be kept at -20 °C (Check!). Exposure of reagents to strong light should be avoided in the process of incubation and storage. All the taps of reagents should be tightened to prevent evaporation and microbial contamination. If not to store reagents according to above suggestions, erroneous results may occur.

Storage: 4 °C/-20 °C

Storage Comment: The unopened kit can be stored at 4℃ for 1 month. If the kit is not used within 1 month, store the items separately according to the conditions since the kit is received.

References for Insulin ELISA Kit (ABIN4993536)

He, Zhao, Xia, Guan, Liu, Wang, Song, Qu, Zuo, Wen, Yu, Huo, Pan, Ban, Zhang, Zhu, Shou, Qiao, Li: "FGF21 ameliorates the neurocontrol of blood pressure in the high fructose-drinking rats." in: Scientific reports, Vol. 6, pp. 29582, (2018) (PubMed).

Durak, Olgar, Tuncay, Karaomerlioglu, Kayki Mutlu, Arioglu Inan, Altan, Turan: "Onset of decreased heart work is correlated with increased heart rate and shortened QT interval in high-carbohydrate fed overweight rats." in: Canadian journal of physiology and pharmacology, Vol. 95, Issue 11, pp. 1335-1342, (2018) (PubMed).

Lembede, Erlwanger, Nkomozepi, Chivandi: "Effect of neonatal orally administered S-allyl cysteine in high-fructose diet fed Wistar rats." in: Journal of developmental origins of health and disease, Vol. 9, Issue 2, pp. 160-171, (2018) (PubMed).

Jin, Ning, Zhou, Wang: "IL-6 Promotes Islet β-Cell Dysfunction in Rat Collagen-Induced Arthritis." in: Journal of diabetes research, Vol. 2016, pp. 7592931, (2017) (PubMed).

Hong, Guo, Wang, Zhao, Xiang, Mei, Abliz, Hu, Deng, Yu: "4-Phenylbutyric Acid Attenuates Pancreatic Beta-Cell Injury in Rats with Experimental Severe Acute Pancreatitis." in: International journal of endocrinology, Vol. 2016, pp. 4592346, (2016) (PubMed).

Yida, Imam, Ismail, Ismail, Ideris, Abdullah: "High fat diet-induced inflammation and oxidative stress are attenuated by N-acetylneuraminic acid in rats." in: Journal of biomedical science, Vol. 22, pp. 96, (2016) (PubMed).

Yida, Imam, Ismail, Ooi, Sarega, Azmi, Ismail, Chan, Hou, Yusuf: "Edible Bird's Nest Prevents High Fat Diet-Induced Insulin Resistance in Rats." in: Journal of diabetes research, Vol. 2015, pp. 760535, (2016) (PubMed).

Target Details for Insulin

Target: 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

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