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IGF1 ELISA Kit

IGF1 Reaktivität: Rind (Kuh) Colorimetric Sandwich ELISA 0.312-20 ng/mL Plasma, Serum
Produktnummer ABIN455447
  • Target Alle IGF1 ELISA Kits anzeigen
    IGF1 (Insulin-Like Growth Factor 1 (IGF1))
    Reaktivität
    • 8
    • 7
    • 6
    • 5
    • 4
    • 3
    • 3
    • 2
    • 2
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    Rind (Kuh)
    Nachweismethode
    Colorimetric
    Methodentyp
    Sandwich ELISA
    Detektionsbereich
    0.312-20 ng/mL
    Untere Nachweisgrenze
    0.312 ng/mL
    Applikation
    ELISA
    Verwendungszweck
    This immunoassay kit allows for the in vitro quantitative determination of bovine IGF-1 concentrations in serum, plasma and other biological fluids.
    Proben
    Plasma, Serum
    Analytische Methode
    Quantitative
    Spezifität
    This assay recognizes recombinant and natural bovine IGF-1.
    Kreuzreaktivität (Details)
    No significant cross-reactivity or interference was observed.
    Sensitivität
    The sensitivity of this assay, or Lower Limit of Detection (LLD) was defined as the lowest detectable concentration that could be differentiated from zero.
    Produktmerkmale
    Bos taurus,Bovine,Insulin-like growth factor I,IGF-I,Somatomedin,IGF1
    Bestandteile
    Reagent (Quantity): Assay plate (1), Standard (2), Sample Diluent (1×20ml), Assay Diluent A (1×10ml), Assay Diluent B (1×10ml), Detection Reagent A (1×120 μl), Detection Reagent B (1×120 μl), Wash Buffer(25 x concentrate) (1×30ml), Substrate (1×10ml), Stop Solution (1×10ml), Plate sealer for 96 wells (5), Instruction (1)
    Benötigtes Material
    Microplate reader. Pipettes and pipette tips. EP tube Deionized or distilled water.
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  • Probenmenge
    100 μL
    Plattentyp
    Pre-coated
    Protokoll
    The microtiter plate provided in this kit has been pre-coated with an antibody specific to IGF-1. Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated polyclonal antibody preparation specific for IGF-1. Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. Then a TMB substrate solution is added to each well. Only those wells that contain IGF-1, biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of a sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450 nm 2 nm. The concentration of IGF-1 in the samples is then determined by comparing the O.D. of the samples to the standard curve.
    Aufbereitung der Reagenzien

    Bring all reagents to room temperature before use. Wash Buffer - If crystals have formed in the concentrate, warm to room temperature and mix gently until the crystals have completely dissolved. Dilute 30 mL of Wash Buffer Concentrate into deionized or distilled water to prepare 750 mL of Wash Buffer. Standard - Reconstitute the Standard with 1.0 ml of Sample Diluent. This reconstitution produces a stock solution of 500 ng/mL. Allow the standard to sit for about 10 minutes with gentle agitation prior to making serial dilutions (Making serial dilution in the wells directly is not permitted). The undiluted standard serves as the highest standard (500 ng/mL). The Sample Diluent serves as the zero standard (0 ng/mL). ng/mL 500 250 125 62.5 31.2 15.6 7.8 0 Detection Reagent A and B - Dilute to the working concentration using Assay Diluent A or B (1:100), respectively.

    Probennahme
    Serum - Use a serum separator tube and allow samples to clot for 30 minutes before centrifugation for 20 minutes at approximately 1000 g. Remove serum and assay immediately or aliquot and store samples at -20 or -80 . Plasma - Collect plasma using EDTA or heparin as an anticoagulant. Centrifuge samples for 15 minutes at 1000 g at 2 - 8 within 30 minutes of collection. Store samples at -20 or -80 . Avoid repeated freeze-thaw cycles. Other biological fluids - Remove particulates by centrifugation and assay immediately or aliquot and store samples at -20 or -80 . Avoid repeated freeze-thaw cycles. Note: Serum and plasma to be used within 7 days may be stored at 2-8 , otherwise samples must stored at -20 ( 1 month) or -80 ( 2 months) to avoid loss of 3 bioactivity and contamination. Avoid freeze-thaw cycles. When performing the assay slowly bring samples to room temperature.
    Testdurchführung

    Allow all reagents to reach room temperature (Please do not dissolve the reagents at 37 directly.). All the reagents should be mixed thoroughly by gently swirling before pipetting. Avoid foaming. Keep appropriate numbers of strips for 1 experiment and 4 remove extra strips from microtiter plate. Removed strips should be resealed and stored at 4 until the kits expiry date. Prepare all reagents, working standards and samples as directed in the previous sections. 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.
    1. Add 100 of Standard, Blank, or Sample per well. Cover with the Plate sealer. Incubate for two hours at 37 .
    2. Remove the liquid of each well, don ’ t wash.
    3. Add 100 μ l of Detection Reagent A working solution to each well. Cover with the Plate sealer. Incubate for 1 hour at 37 . Detection Reagent A working solution may appear cloudy. Warm to room temperature and mix gently until solution appears uniform.
    4. Aspirate each well and wash, repeating the process three times for a total of three washes. Wash by filling each well with Wash Buffer (approximately 400 μ l) using a squirt bottle, multi-channel pipette, manifold dispenser 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.
    5. Add 100 μ l of Detection Reagent B working solution to each well. Cover with a new Plate sealer. Incubate for one hour at 37 .
    6. Repeat the aspiration/wash process for five times as conducted in step
    4. 7. Add 90 μ l of Substrate Solution to each well. Cover with a new Plate sealer. Incubate for 15 - 30 minutes at 37 . Protect from light.
    8. Add 50 μ l of Stop Solution to each well. If color change does not appear uniform, gently tap the plate to ensure thorough mixing.
    9. Determine the optical density of each well at once, using a microplate reader set to 450 nm.
    Important Note:
    1. Absorbance is a function of the incubation time. Therefore, prior to starting the assay it is recommended that all reagents should be freshly prepared prior to use and all required strip-wells are secured in the microtiter frame. This will ensure equal elapsed time for each pipetting step, without interruption.
    2. Please carefully reconstitute Standards or working Detection Reagent A and B according to the instruction, and avoid foaming and mix gently until the crystals have completely dissolved. The reconstituted Standards can be used only once. This assay requires pipetting of small volumes. To minimize imprecision caused by pipetting, ensure that pipettors are calibrated. It is recommended to suck more than 10 μ l for once pipetting.
    3. To ensure accurate results, proper adhesion of plate sealers during incubation steps is necessary. Do not allow wells to sit uncovered for extended periods between incubation steps. Once reagents have been added to the well strips, DO NOT let the strips DRY at any time during the assay. 5
    4. For each step in the procedure, total dispensing time for addition of reagents to the assay plate should not exceed 10 minutes.
    5. To avoid cross-contamination, change pipette tips between additions of each standard level, between sample additions, and between reagent additions. Also, use separate reservoirs for each reagent.
    6. The wash procedure is critical. Insufficient washing will result in poor precision and falsely elevated absorbance readings.
    7. Duplication of all standards and specimens, although not required, is recommended.
    8. Substrate Solution is easily contaminated. Please protect it from light.

    Ergebnisberechnung

    Average the duplicate readings for each standard, control, and samples and subtract the average zero standard optical density. Create a standard curve by reducing the data using computer software capable of generating a four parameter logistic (4-PL) curve-fit. As an alternative, construct a standard curve by plotting the mean absorbance for each standard on the x-axis against the concentration on the y-axis and draw a best fit curve through the points on the graph. The data may be linearized by plotting the log of the IGF-1 concentrations versus the log of the O.D. and the best fit line can be determined by regression analysis. It is recommended to use some related software to do this calculation, such as curve expert 13.0. This procedure will produce an adequate but less precise fit of the data. If samples have been diluted, the concentration read from the standard curve must be multiplied by the dilution factor.

    Beschränkungen
    Nur für Forschungszwecke einsetzbar
  • Handhabung
    1. The kit should not be used beyond the expiration date on the kit label.
    2. Do not mix or substitute reagents with those from other lots or sources.
    3. If samples generate values higher than the highest standard, further dilute the samples and repeat the assay. Any variation in standard diluent, operator, pipetting technique, washing technique,incubation time or temperature, and kit age can cause variation in binding.
    4. This assay is designed to eliminate interference by soluble receptors, ligands, binding proteins, and other factors present in biological samples. Until all factors have been tested in the Immunoassay, the possibility of interference cannot be excluded.
    5. Limited by the current condition and scientific technology, we can't 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.
    Lagerung
    4 °C/-20 °C
    Informationen zur Lagerung
    The Standard, Detection Reagent A, Detection Reagent B and the 96-well strip plate should be stored at -20 °C upon being received. The other reagents can be stored at 4 °C.
  • Target Alle IGF1 ELISA Kits anzeigen
    IGF1 (Insulin-Like Growth Factor 1 (IGF1))
    Andere Bezeichnung
    IGF1 (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
    Insulin-like Growth Factor I (IGF-I), also known as somatomedin C, is a member of the insulin superfamily. It was originally discovered as a mediator of growth hormone actions on somatic cell growth, but has also been shown to be an important regulator of cell metabolism, differentiation and survival. IGF-I is synthesized as a preproprotein that is proteolytically cleaved to generate the mature protein linked by three disulfide bonds. Mature IGF-I is highly conserved among mammals, with 100% sequence identity between the Porcine, bovine, porcine, equine and canine proteins. IGF-I is synthesized in the liver and multiple other tissues. It is found in blood and other body fluids as a complex with specific high affinity IGF binding proteins (IGFBP-1to-6). The IGFBPs are expressed in specific patterns during development. They are modulators of IGF actions, which control IGF bioavailability to specific cell-surface receptors. Their functions are further regulated by IGFBP proteases, which proteolytically cleave the IGFBPs to lower the affinity with which they bind IGFs and increase IGF bioavailability. Some IGFBPs also have IGF-independent effects on cell functions. IGF-I circulates primarily as a ternary complex with IGFBP-3 or IGFBP-5 and the acid-labile subunit (ALS). Some IGF-I is also present in binary complexes with other IGFBPs. Whereas the ternary complexes are generally restricted to the vasculature, the binary complexes freely enter the tissues. IGF-I actions are mediated by two type I transmembrane receptor tyrosine kinases: the IGF-I receptor (IGF-I R), and the insulin receptor (INS R) that exists in two alternatively spliced isoforms (INS R-A and -B). Both IGF-I R and INS R share a highly homologous structure and are ubiquitously expressed. Functional IGF-I receptors are tetrameric glycoproteins composed of two disulfide-linked IGF-I Rs or disulfide-linked hybrids of one IGF-I R and one INS R. Whereas IGF-I binds with high-affinity to homodimeric IGF-I R and heterdimeric IGF-I R:INS R-A or – B hybrids, high-affinity binding of insulin is observed only with dimeric INS R or IGF-I R:INS R-A hybrid but not with IGF-I R:INS R-B hybrid. The signaling responses from the various receptors are different depending whether insulin or IGF-I is used as the activating ligand.
    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
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