IGF1 ELISA Kit

Produktnummer ABIN625003

Produktdetails IGF1 ELISA Kit

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

Reaktivität: Human

Nachweismethode: Colorimetric

Methodentyp: Sandwich ELISA

Detektionsbereich: 0.1-30 ng/mL

Untere Nachweisgrenze: 0.1 ng/mL

Applikation: ELISA

Verwendungszweck: Human IGF-1 ELISA Kit for cell culture supernatants, plasma, and serum samples.

Proben: Plasma, Cell Culture Supernatant, Serum

Analytische Methode: Quantitative

Spezifität: This IGF-1 ELISA kit shows no cross-reactivity with any of the cytokines tested: Human Angiogenin, BDNF, BLC, ENA-78, FGF-4, IGFBP-1, IGFBP-2, IGFBP-3, IGFBP-4, IL-1 alpha, IL-1 beta, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12 p70, IL-12 p40, IL-13, IL-15, I-309, IP-10, G-CSF, GM-CSF, IFN-gamma, Leptin (OB), MCP-1, MCP-3, MDC, MIP-1 alpha, MIP-1 beta, MIP-1 delta, PARC, RANTES, SCF, TARC, TGF-beta, TIMP-1, TIMP-2, TNF-alpha, TNF-beta, TPO, VEGF.

Sensitivität: 100 pg/mL

Produktmerkmale:

• Strip plates and additional reagents allow for use in multiple experiments
• Quantitative protein detection
• Establishes normal range
• The best products for confirmation of antibody array data

Bestandteile:

• Pre-Coated 96-well Strip Microplate
• Wash Buffer
• Stop Solution
• Assay Diluent(s)
• Lyophilized Standard
• Biotinylated Detection Antibody
• Streptavidin-Conjugated HRP
• TMB One-Step Substrate

Benötigtes Material:

• Distilled or deionized water
• Precision pipettes to deliver 2 μL to 1 μL volumes
• Adjustable 1-25 μL pipettes for reagent preparation
• 100 μL and 1 liter graduated cylinders
• Tubes to prepare standard and sample dilutions
• Absorbent paper
• Microplate reader capable of measuring absorbance at 450nm
• Log-log graph paper or computer and software for ELISA data analysis

Anwendungsinformationen

Applikationshinweise: Recommended Dilution for serum and plasma samples2 - 20 fold

Probenmenge: 100 μL

Plattentyp: Pre-coated

Protokoll:

1. Prepare all reagents, samples and standards as instructed in the manual.
2. Add 100 μL of standard or sample to each well.
3. Incubate 2.5 h at RT or O/N at 4 °C.
4. Add 100 μL of prepared biotin antibody to each well.
5. Incubate 1 h at RT.
6. Add 100 μL of prepared Streptavidin solution to each well.
7. Incubate 45 min at RT.
8. Add 100 μL of TMB One-Step Substrate Reagent to each well.
9. Incubate 30 min at RT.
10. Add 50 μL of Stop Solution to each well.
11. Read at 450 nm immediately.

Aufbereitung der Reagenzien:

1. Bring all reagents and samples to room temperature (18 - 25 °C) before use.
   2. Sample dilution: If your samples need to be diluted, Assay Diluent C (Item L) should be used for dilution of serum/plasma/culture supernatants/urine. Suggested dilution for normal serum/plasma: 2-20 fold. Please note that levels of the target protein may vary between different specimens. Optimal dilution factors for each sample must be determined by the investigator.
   3. Preparation of standard: Briefly spin the vial of Item C and then add 400 µL Assay Diluent C into Item C vial to prepare a 100 ng/mL standard. Dissolve the powder thoroughly by a gentle mix. Add 150 µL IGF-I standard from the vial of Item C, into a tube with 350 µL Assay Diluent C to prepare a 30 ng/mL standard solution. Pipette 300 µL Assay Diluent C into each tube. Use the 30 ng/mL standard solution to produce a dilution series . Mix each tube thoroughly before the next transfer. Assay Diluent C serves as the zero standard (0 ng/mL). 200 µL 200myl 200 µL 200 µL 200 µL 200 µL 150 µL Standard + 350 µL 30 12 4.8 1.92 0.768 0.307 0.123 0 ng/mL ng/mL ng/mL ng/mL ng/mL ng/mL ng/mL ng/mL
   4. If the Wash Concentrate (20x) (Item B) contains visible crystals, warm to room temperature and mix gently until dissolved. Dilute 20 ml of Wash Buffer Concentrate into deionized or distilled water to yield 400 ml of 1x Wash Buffer.
   5. Briefly spin the Detection Antibody vial (Item F) before use. Add 100 µL of Assay Diluent C into the vial to prepare a detection antibody concentrate. Pipette up and down to mix gently (the concentrate can be stored at 4 °C for 5 days). The detection antibody concentrate should be diluted 80-fold with Assay Diluent C and used in step 4 of Part VI Assay Procedure.
   6. Briefly spin the HRP-Streptavidin concentrate vial (Item G) before use. HRP-Streptavidin concentrate should be diluted 120-fold with Assay Diluent C. For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 100 µL of HRP-Streptavidin concentrate into a tube with 12 ml Assay Diluent C to prepare a final 120 fold diluted HRP- Streptavidin solution (don't store the diluted solution for next day use). Mix well.

Testdurchführung:

1. Bring all reagents and samples to room temperature (18 - 25 °C) before use. It is recommended that all standards and samples be run at least in duplicate.
   2. Add 100 µL of each standard (see Reagent Preparation step 2) and sample into appropriate wells. Cover well and incubate for 2.5 hours at room temperature or over night at 4 °C with gentle shaking.
   3. Discard the solution and wash 4 times with 1x Wash Solution. Wash by filling each well with Wash Buffer (300 myl) using a multi-channel Pipette or autowasher. Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Invert the plate and blot it against clean paper towels.
   4. Add 100 µL of 1x prepared biotinylated antibody (Reagent Preparation step 5) to each well. Incubate for 1 hour at room temperature with gentle shaking.
   5. Discard the solution. Repeat the wash as in step
   6. Add 100 µL of prepared Streptavidin solution (see Reagent Preparation step 6) to each well. Incubate for 45 minutes at room temperature with gentle shaking.
   7. Discard the solution. Repeat the wash as in step
   8. Add 100 µL of TMB One-Step Substrate Reagent (Item H) to each well. Incubate for 30 minutes at room temperature in the dark with gentle shaking.
   9. Add 50 µL of Stop Solution (Item I) to each well. Read at 450 nm immediately.

Ergebnisberechnung:

Calculate the mean absorbance for each set of duplicate standards, controls and samples, and subtract the average zero standard optical density. Plot the standard curve on log-log graph paper or using Sigma plot software, with standard concentration on the x-axis and absorbance on the y-axis. Draw the best-fit straight line through the standard points.
Typical Data: These standard curves are for demonstration only. A standard curve must be run with each assay. Assay Diluent C Human IGF-I concentration (ng/mL) O D =4 50 n m 0.01 0.1 1 10 0.01 0.1 1 10 100
Sensitivity: The minimum detectable dose of IGF-1 is typically less than 0.1 ng/mL.
Recovery: Recovery was determined by spiking various levels of IGF-1 into human serum, plasma and cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Serum 119.6 105-138 Plasma 105.4 81-121 Cell culture media 102.5 98-110
Linearity: Sample Type Serum Plasma Cell Culture Media 1:2 Average % of Expected 116.4 111.9 103.3 Range ( %) 108-124 107-125 94-118 1:4 Average % of Expected 121.9 104.6 84.62 Range ( %) 114-130 81-120 77-92
Reproducibility: Intra-Assay: CV<10 % Inter-Assay: CV<12 %

Testpräzision: Intra-Assay: CV< 10 % Inter-Assay: CV< 12 %

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Handhabung: Avoid repeated freeze-thaw cycles.

Lagerung: -20 °C

Informationen zur Lagerung: The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.

Haltbarkeit: 6 months

Referenzen für IGF1 ELISA Kit (ABIN625003)

Shen, Lie, Miao, Yu, Lu, Feng, Li, Zu, Liu, Li: "Conditioned medium from umbilical cord mesenchymal stem cells induces migration and angiogenesis." in: Molecular medicine reports, Vol. 12, Issue 1, pp. 20-30, (2015) (PubMed).

Pickard, McDade, McFarland, McCluggage, Wheeler, McCance: "HPV16 Down-Regulates the Insulin-Like Growth Factor Binding Protein 2 to Promote Epithelial Invasion in Organotypic Cultures." in: PLoS pathogens, Vol. 11, Issue 6, pp. e1004988, (2015) (PubMed).

Meeks, Van Haitsma, Mast, Arnold, Streim, Sephton, Smith, Kleban, Rovine: "Psychological and social resources relate to biomarkers of allostasis in newly admitted nursing home residents." in: Aging & mental health, Vol. 20, Issue 1, pp. 88-99, (2015) (PubMed).

Stoppoloni, Politi, Leopizzi, Gaetani, Guazzo, Basciani, Moreschini, De Santi, Scandurra, Scotto dAbusco: "Effect of glucosamine and its peptidyl-derivative on the production of extracellular matrix components by human primary chondrocytes." in: Osteoarthritis and cartilage / OARS, Osteoarthritis Research Society, Vol. 23, Issue 1, pp. 103-13, (2014) (PubMed).

Ritsche, Nindl, Wideman: "Exercise-Induced growth hormone during acute sleep deprivation." in: Physiological reports, Vol. 2, Issue 10, (2014) (PubMed).

Krogh, Rostrup, Thomsen, Elfving, Videbech, Nordentoft: "The effect of exercise on hippocampal volume and neurotrophines in patients with major depression--a randomized clinical trial." in: Journal of affective disorders, Vol. 165, pp. 24-30, (2014) (PubMed).

Aucouturier, Thivel, Isacco, Fellmann, Chardigny, Duclos, Duché: "Combined food intake and exercise unmask different hormonal responses in lean and obese children." in: Applied physiology, nutrition, and metabolism = Physiologie appliquée, nutrition et métabolisme, Vol. 38, Issue 6, pp. 638-43, (2013) (PubMed).

De Barros, Dehez, Arnaud, Barreau, Cazavet, Perez, Galinier, Casteilla, Planat-Bénard: "Aging-related decrease of human ASC angiogenic potential is reversed by hypoxia preconditioning through ROS production." in: Molecular therapy : the journal of the American Society of Gene Therapy, Vol. 21, Issue 2, pp. 399-408, (2013) (PubMed).

Kim, Kang, Krueger, Sen, Holcomb, Chen, Wenke, Yang: "Sequential delivery of BMP-2 and IGF-1 using a chitosan gel with gelatin microspheres enhances early osteoblastic differentiation." in: Acta biomaterialia, Vol. 8, Issue 5, pp. 1768-77, (2012) (PubMed).

Nwozo, Oyinloye: "Hepatoprotective effect of aqueous extract of Aframomum melegueta on ethanol-induced toxicity in rats." in: Acta biochimica Polonica, Vol. 58, Issue 3, pp. 355-8, (2012) (PubMed).

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Details zu IGF1

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

Andere Bezeichnung: IGF-I (IGF1 Produkte)

Synonyme: IGF-I ELISA Kit, IGF1A ELISA Kit, IGFI ELISA Kit, C730016P09Rik ELISA Kit, Igf-1 ELISA Kit, Igf-I ELISA Kit, Npt2B ELISA Kit, IGF-1 ELISA Kit, IGFIA ELISA Kit, IGF-IB ELISA Kit, igf1 ELISA Kit, IGF-1L ELISA Kit, IGF-1a ELISA Kit, IGF1 ELISA Kit, igf-1 ELISA Kit, igf1-A ELISA Kit, igf1.S ELISA Kit, xigf1 ELISA Kit, insulin like growth factor 1 ELISA Kit, insulin-like growth factor 1 ELISA Kit, insulin-like growth factor I ELISA Kit, insulin like growth factor 1 L homeolog ELISA Kit, IGF1 ELISA Kit, Igf1 ELISA Kit, LOC100136741 ELISA Kit, igf1 ELISA Kit, LOC678666 ELISA Kit, igf1.L ELISA Kit, LOC104911603 ELISA Kit

Hintergrund: The Human IGF-I ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of human IGF-I in serum, plasma, cell culture supernatants and urine. This assay employs an antibody specific for human IGF-I coated on a 96-well plate. Standards and samples are pipetted into the wells and IGF-I present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-human IGF-I antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of IGF-I bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm. Reproducibility: Intra-Assay: CV<10% Inter-Assay: CV<12%.

Gen-ID: 3479

UniProt: P05019

Pathways: RTK Signalweg, 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, Autophagie, Smooth Muscle Cell Migration, Activated T Cell Proliferation, Positive Regulation of fat Cell Differentiation