IGF1 ELISA Kit

Catalog No. ABIN6956789

Product Details IGF1 ELISA Kit

Target: IGF1 (Insulin-Like Growth Factor 1 (IGF1))

Reactivity: Rat

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 0.15 ng/mL - 10 ng/mL

Minimum Detection Limit: 0.15 ng/mL

Application: ELISA

Purpose: The kit is a sandwich enzyme immunoassay for in vitro quantitative measurement of IGF1 in rat serum, plasma, tissue homogenates, cell lysates, cell culture supernates.

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 Like Growth Factor 1 (IGF1)

Sensitivity: 0.069 ng/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

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: 100 μL

Assay Time: 3 h

Plate: Pre-coated

Protocol:

1. Prepare all reagents, samples and standards,
2. Add 100μL standard or sample to each well. Incubate 1 hours at 37 °C,
3. Aspirate and add 100μL prepared Detection Reagent A. Incubate 1 hour at 37 °C,
4. Aspirate and wash 3 times,
5. Add 100μL prepared Detection Reagent B. Incubate 30 minutes at 37 °C,
6. Aspirate and wash 5 times,
7. Add 90μL Substrate Solution. Incubate 10-20 minutes at 37 °C,
8. Add 50μL Stop Solution. Read at 450nm 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 1.0 mL of Standard Diluent, keep for 10 minutes at room temperature, shake gently (not to foam). The concentration of the standard in the stock solution is 20 ng/mL. Firstly dilute the stock solution to 10 ng/mL and the diluted standard serves as the highest standard (10 ng/mL). Then prepare 7 tubes containing 0.5 mL Standard Diluent and use the diluted standard to produce a double dilution series. Mix each tube thoroughly before the next transfer. Set up 7 points of diluted standard such as 10 ng/mL, 5 ng/mL, 2.5 ng/mL, 1.25 ng/mL, 0.625 ng/mL, 0.312 ng/mL, 0.156 ng/mL, and the last microcentrifuge tube with Standard Diluent is the blank as 0 ng/mL.
3. Detection Reagent A and Detection Reagent B - If lyophilized reconstitute the Detection Reagent A with 150μL of Reagent Diluent, keep 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 standards within 15 minutes before assay. Please do not dissolve the reagents at 37 °C directly.
3. 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.
4. The reconstituted Standards, Detection Reagent A and Detection Reagent B can be used only once.
5. If crystals have formed in the Wash Solution concentrate (30x), warm to room temperature and mix gently until the crystals are completely dissolved.
6. 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

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.

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.

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Expiry Date: 6 months

References for IGF1 ELISA Kit (ABIN6956789)

Wang, Hai, Niu, Ai, Cao, Li, Li: "Chronic intermittent hypoxia disturbs insulin secretion and causes pancreatic injury via the MAPK signaling pathway." in: Biochemistry and cell biology = Biochimie et biologie cellulaire, Vol. 95, Issue 3, pp. 415-420, (2017) (PubMed).

de Almeida, Gomes da Silva, Lopim, Vannucci Campos, Fernandes, Cabral, Arida: "Resistance Exercise Reduces Seizure Occurrence, Attenuates Memory Deficits and Restores BDNF Signaling in Rats with Chronic Epilepsy." in: Neurochemical research, Vol. 42, Issue 4, pp. 1230-1239, (2017) (PubMed).

Fernandes, Soares, do Amaral Baliego, Arida: "A single bout of resistance exercise improves memory consolidation and increases the expression of synaptic proteins in the hippocampus." in: Hippocampus, Vol. 26, Issue 8, pp. 1096-103, (2016) (PubMed).

Kose, Bastug, Sonmez, Per, Ozdemir, Kaymak, Yahşi, Ozturk: "Protective effect of vitamin D against hyperoxia-induced lung injury in newborn rats." in: Pediatric pulmonology, (2016) (PubMed).

Giampá, Mônico-Neto, de Mello, Souza, Tufik, Lee, Koike, Dos Santos, Antonio, Serra, Tucci, Antunes: "Paradoxical Sleep Deprivation Causes Cardiac Dysfunction and the Impairment Is Attenuated by Resistance Training." in: PLoS ONE, Vol. 11, Issue 11, pp. e0167029, (2016) (PubMed).

Sangun, Dundar, Darici, Comlekci, Doguc, Celik: "The effects of long-term exposure to a 2450?MHz electromagnetic field on growth and pubertal development in female Wistar rats." in: Electromagnetic biology and medicine, Vol. 34, Issue 1, pp. 63-71, (2015) (PubMed).

Mônico-Neto, Giampá, Lee, de Melo, Souza, Dáttilo, Minali, Santos Prado, Tufik, de Mello, Antunes: "Negative energy balance induced by paradoxical sleep deprivation causes multicompartmental changes in adipose tissue and skeletal muscle." in: International journal of endocrinology, Vol. 2015, pp. 908159, (2015) (PubMed).

Ergenoglu, Yildirim, Yildirim, Yeniel, Erbas, Yavasoglu, Taskiran, Karadadas: "Effects of Resveratrol on Ovarian Morphology, Plasma Anti-Mullerian Hormone, IGF-1 Levels, and Oxidative Stress Parameters in a Rat Model of Polycystic Ovary Syndrome." in: Reproductive sciences (Thousand Oaks, Calif.), Vol. 22, Issue 8, pp. 942-7, (2015) (PubMed).

Shi, Huang, Wang, Huang: "The Protective Effects of Exclusive Enteral Nutrition Formulas on Growth Factor Expression and the Proximal Tibial Epiphyseal Growth Plate in a TNBS-Induced IBD Rat Model." in: Digestive diseases and sciences, Vol. 60, Issue 7, pp. 1931-40, (2015) (PubMed).

Song, Yang, Tang, Chen: "Effects of sericin on the testicular growth hormone/insulin-like growth factor-1 axis in a rat model of type 2 diabetes." in: International journal of clinical and experimental medicine, Vol. 8, Issue 7, pp. 10411-9, (2015) (PubMed).

Guo, Gao, Lin, Wu, Huang, Dong, Chen, Lu, Fu, Wang, Ma, Chen, Wu, He, Yang, Liao, Zheng, Tan: "BMSCs reduce rat granulosa cell apoptosis induced by cisplatin and perimenopause." in: BMC cell biology, Vol. 14, Issue 1, pp. 18, (2014) (PubMed).

Simko, Fekete, Gradosova, Malakova, Zivna, Valis, Palicka, Zivny: "The effect of topiramate and lamotrigine on rat bone mass, structure and metabolism." in: Journal of the neurological sciences, Vol. 340, Issue 1-2, pp. 80-5, (2014) (PubMed).

Haleagrahara, Chakravarthi, Mathews: "Insulin like growth factor-1 (IGF-1) causes overproduction of IL-8, an angiogenic cytokine and stimulates neovascularization in isoproterenol-induced myocardial infarction in rats." in: International journal of molecular sciences, Vol. 12, Issue 12, pp. 8562-74, (2012) (PubMed).

Gradosova, Zivna, Palicka, Hubena, Svejkovska, Zivny: "Protective effect of amlodipine on rat bone tissue after orchidectomy." in: Pharmacology, Vol. 89, Issue 1-2, pp. 37-43, (2012) (PubMed).

Lisa, Haleagrahara, Chakravarthi: "Insulin-Like Growth Factor-1 (IGF-1) Reduces ischemic changes and increases circulating angiogenic factors in experimentally - induced myocardial infarction in rats." in: Vascular cell, Vol. 3, Issue 1, pp. 13, (2011) (PubMed).

Al-Massadi, Trujillo, Señaris, Pardo, Castelao, Casanueva, Seoane: "The vagus nerve as a regulator of growth hormone secretion." in: Regulatory peptides, Vol. 166, Issue 1-3, pp. 3-8, (2010) (PubMed).

Qin, Tian: "A microarray gene analysis of peripheral whole blood in normal adult male rats after long-term GH gene therapy." in: Cellular & molecular biology letters, Vol. 15, Issue 2, pp. 177-95, (2010) (PubMed).

Qin, Tian: "Characterization of the specific and sustained GH1 expression induced by rAAV2/1 in normal adult male rats." in: Molecular biology reports, Vol. 37, Issue 7, pp. 3643-51, (2010) (PubMed).

Gozal, Nair, Goldbart: "Physical activity attenuates intermittent hypoxia-induced spatial learning deficits and oxidative stress." in: American journal of respiratory and critical care medicine, Vol. 182, Issue 1, pp. 104-12, (2010) (PubMed).

Taqi, Wallace, de Heuvel, Chelikani, Zheng, Berthoud, Holst, Sigalet: "The influence of nutrients, biliary-pancreatic secretions, and systemic trophic hormones on intestinal adaptation in a Roux-en-Y bypass model." in: Journal of pediatric surgery, Vol. 45, Issue 5, pp. 987-95, (2010) (PubMed).

Target Details for IGF1

Target: IGF1 (Insulin-Like Growth Factor 1 (IGF1))

Alternative Name: Insulin Like Growth Factor 1 (IGF1) (IGF1 Products)

Pathways: RTK Signaling, Intracellular Steroid Hormone Receptor Signaling Pathway, Peptide Hormone Metabolism, Hormone Activity, Regulation of Intracellular Steroid Hormone Receptor Signaling, Regulation of Hormone Metabolic Process, Regulation of Hormone Biosynthetic Process, Stem Cell Maintenance, Glycosaminoglycan Metabolic Process, Regulation of Carbohydrate Metabolic Process, Autophagy, Smooth Muscle Cell Migration, Activated T Cell Proliferation, Positive Regulation of fat Cell Differentiation