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IGFBPI ELISA Kit

IGFBPI Reaktivität: Human Colorimetric Sandwich ELISA 5-2000 pg/mL Cell Culture Supernatant, Plasma, Serum
Produktnummer ABIN1979697
  • Target Alle IGFBPI ELISA Kits anzeigen
    IGFBPI (Insulin-Like Growth Factor Binding Protein 1 (IGFBPI))
    Reaktivität
    • 6
    • 6
    • 3
    • 3
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    Human
    Nachweismethode
    Colorimetric
    Methodentyp
    Sandwich ELISA
    Detektionsbereich
    5-2000 pg/mL
    Untere Nachweisgrenze
    5 pg/mL
    Applikation
    ELISA
    Verwendungszweck
    Human IGFBP-1 ELISA Kit for cell culture supernatants, plasma, and serum samples.
    Proben
    Plasma, Cell Culture Supernatant, Serum
    Analytische Methode
    Quantitative
    Spezifität
    This ELISA kit shows no cross-reactivity with any of the cytokines tested: Human BDNF, BLC, ENA-78, FGF-4, IL-1 alpha, IL-1 beta, IL-2, IL-3, IL-4, IL-5, 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, IGFBP-2, IGFBP-3, IGFBP-4, Leptin (OB), MCP-1, MCP-2, MCP-3, MDC, MIP-1 alpha, MIP-1 beta, MIP-1 delta, PARC, PDGF, RANTES, SCF, TARC, TGF-beta, TIMP-1, TIMP-2, TNF-alpha, TNF-beta, TPO, VEGF.
    Kreuzreaktivität (Details)
    This ELISA kit shows no cross-reactivity with any of the cytokines tested (e.g., human BDNF, BLC, ENA-78, FGF-4, IL-1alpha, IL-1beta, IL-2, IL-3, IL-4, IL-5, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12 p70, IL-12 p40, IL-13, IL-15, IL-309, IP-10, G-CSF, GM-CSF, IFN-gamma, IGFBP-2, IGFBP-3, IGFBP-4, Leptin (OB), MCP-1, MCP-2, MCP-3, MDC, MIP-1alpha, MIP-1 beta, MIP-1delta, PARC, PDGF, RANTES, SCF, TARC, TGF-beta, TIMP-1, TIMP-2, TNF-alpha, TNF-beta, TPO, VEGF).
    Sensitivität
    < 5 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
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  • Applikationshinweise
    Recommended Dilution for serum and plasma samples5 - 500 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 A (Item D) should be used for dilution of serum/plasma samples. 1x Assay Diluent B (Item E) should be used for dilution of culture supernatants and urine. Suggested dilution for normal serum/plasma: 5-500 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. Assay Diluent B should be diluted 5-fold with deionized or distilled water.
      4. Preparation of standard: Briefly spin the vial of Item C and then add 400 µL Assay Diluent A (for serum/plasma samples) or 1x Assay Diluent B (for cell culture medium and urine) into Item C vial to prepare a 50 ng/mL standard. Dissolve the powder thoroughly by a gentle mix. Add 40 µL IGF-BP-1 standard from the vial of Item C, into a tube with 960 µL Assay Diluent A or 1x Assay Diluent B to prepare a 2000 pg/mL stock standard solution. Pipette 400 µL Assay Diluent A or 1x Assay Diluent B into each tube. Use the stock standard solution to produce a dilution series . Mix each tube thoroughly before the next transfer. Assay Diluent A or 1x Assay Diluent B serves as the zero standard (0 pg/mL). 200 µL 40 µL standard + 960 µL 200myl 200 µL 200 µL 200 µL 200 µL 2000 666.7 222.2 74.07 24.69 8.23 2.74 0 pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL pg/mL
      5. 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.
      6. Briefly spin the Detection Antibody vial (Item F) before use. Add 100 µL of 1x Assay Diluent B 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 1x Assay Diluent B and used in step 4 of Part VI Assay Procedure.
      7. Briefly spin the HRP-Streptavidin concentrate vial (Item G) and pipette up and down to mix gently before use. HRP-Streptavidin concentrate should be diluted 240-fold with 1x Assay Diluent B. For example: Briefly spin the vial (Item G) and pipette up and down to mix gently . Add 50 µL of HRP-Streptavidin concentrate into a tube with 12 ml 1x Assay Diluent B to prepare a final 240 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 6) 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 7) 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 A Human IGF-BP-1 concentration (pg/mL) O D =4 50 (n m ) 0.001 0.01 0.1 1 10 1 10 100 1,000 10,000 Assay Diluent B Human IGF-BP-1 concentration (pg/mL) O D =4 50 (n m ) 0.01 0.1 1 10 1 10 100 1,000 10,000
    Sensitivity: The minimum detectable dose of IGF-BP-1 is typically less than 5 pg/mL.
    Recovery: Recovery was determined by spiking various levels of human IGF-BP-1 into human serum, plasma and cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Serum 95.27 84-104 Plasma 92.49 82-102 Cell culture media 94.64 83-102
    Linearity: Sample Type Serum Plasma Cell culture media 1:2 Average % of Expected 93 92 94 Range ( %) 81-101 82-102 83-103 1:4 Average % of Expected 94 93 93 Range ( %) 83-102 84-103 82-102
    Reproducibility: Intra-Assay: CV<10 % Inter-Assay: CV<12 %

    Testpräzision
    Intra-Assay: CV< 10 % Inter-Assay: CV< 12 %
    Beschränkungen
    Nur für Forschungszwecke einsetzbar
  • 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
  • Pineda, Lu, Cao, Kim: "Influence of Cancer-Associated Endometrial Stromal Cells on Hormone-Driven Endometrial Tumor Growth." in: Hormones & cancer, Vol. 6, Issue 4, pp. 131-41, (2015) (PubMed).

    Founds, Zeng, Lykins, Roberts: "Developing Potential Candidates of Preclinical Preeclampsia." in: International journal of molecular sciences, Vol. 16, Issue 11, pp. 27208-27, (2015) (PubMed).

    Pflum, Palumbo, Li: "Adverse effect of demineralized bone powder on osteogenesis of human mesenchymal stem cells." in: Experimental cell research, Vol. 319, Issue 13, pp. 1942-55, (2013) (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).

    Estella, Herrer, Moreno-Moya, Quiñonero, Martínez, Pellicer, Simón: "miRNA signature and Dicer requirement during human endometrial stromal decidualization in vitro." in: PLoS ONE, Vol. 7, Issue 7, pp. e41080, (2012) (PubMed).

    Ammoun, Schmid, Zhou, Ristic, Ercolano, Hilton, Perks, Hanemann: "Insulin-like growth factor-binding protein-1 (IGFBP-1) regulates human schwannoma proliferation, adhesion and survival." in: Oncogene, Vol. 31, Issue 13, pp. 1710-22, (2012) (PubMed).

    Pereira-Fantini, Thomas, Taylor, Nagy, Sourial, Fuller, Bines: "Colostrum supplementation restores insulin-like growth factor -1 levels and alters muscle morphology following massive small bowel resection." in: JPEN. Journal of parenteral and enteral nutrition, Vol. 32, Issue 3, pp. 266-75, (2008) (PubMed).

    Angervo: "Epidermal growth factor enhances insulin-like growth factor binding protein-1 synthesis in human hepatoma cells." in: Biochemical and biophysical research communications, Vol. 189, Issue 2, pp. 1177-83, (1993) (PubMed).

    Kratz, Lake, Ljungström, Forsberg, Haegerstrand, Gidlund: "Effect of recombinant IGF binding protein-1 on primary cultures of human keratinocytes and fibroblasts: selective enhancement of IGF-1 but not IGF-2-induced cell proliferation." in: Experimental cell research, Vol. 202, Issue 2, pp. 381-5, (1992) (PubMed).

    Frost, Tseng: "Insulin-like growth factor-binding protein-1 is phosphorylated by cultured human endometrial stromal cells and multiple protein kinases in vitro." in: The Journal of biological chemistry, Vol. 266, Issue 27, pp. 18082-8, (1991) (PubMed).

  • Target Alle IGFBPI ELISA Kits anzeigen
    IGFBPI (Insulin-Like Growth Factor Binding Protein 1 (IGFBPI))
    Andere Bezeichnung
    IGFBP-1 (IGFBPI Produkte)
    Synonyme
    cb656 ELISA Kit, igfbp1 ELISA Kit, IGFBP-1 ELISA Kit, MGC68538 ELISA Kit, afbp ELISA Kit, ibp1 ELISA Kit, pp12 ELISA Kit, igf-bp25 ELISA Kit, higfbp-1 ELISA Kit, IGFBP1 ELISA Kit, LOC100223208 ELISA Kit, LOC100305121 ELISA Kit, AFBP ELISA Kit, IBP1 ELISA Kit, IGF-BP25 ELISA Kit, PP12 ELISA Kit, hIGFBP-1 ELISA Kit, IGFBA ELISA Kit, insulin like growth factor binding protein 1 ELISA Kit, insulin-like growth factor binding protein 1a ELISA Kit, insulin like growth factor binding protein 1 S homeolog ELISA Kit, insulin like growth factor binding protein 1 L homeolog ELISA Kit, insulin-like growth factor binding protein 1 ELISA Kit, IGFBP1 ELISA Kit, igfbp1a ELISA Kit, igfbp1.S ELISA Kit, igfbp1.L ELISA Kit, igfbp1 ELISA Kit, LOC100305121 ELISA Kit, Igfbp1 ELISA Kit
    Hintergrund
    IGF-BPs (Insulin-like growth factor binding proteins) are found in various body fluids such as blood serum, amniotic fluid, and liquor. They are synthesized in the liver and are produced also by various tumor cell lines and cell types. IGF-BP-1 is found predominantly in the placenta and the amniotic fluid. It has been shown also to be an inhibitor of IGF mitogenic activities for human breast cancer cells. The effect of IGF-BP-1 depends on its phosphorylation status, phosphorylated IGF-BP-1 inhibits IGF actions whereas the nonphosphorylated isoform is stimulatory. Predominant sites of IGF-BP-1 transcription in the human fetal kidney are those with most active differentiation.
    Gen-ID
    3484
    UniProt
    P08833
    Pathways
    Myometrial Relaxation and Contraction, ER-Nucleus Signaling, Growth Factor Binding
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