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G-CSF ELISA 试剂盒

CSF3 适用: 人 Colorimetric Sandwich ELISA 1-500 pg/mL Cell Culture Supernatant, Plasma, Serum
产品编号 ABIN624983
发货至: 中国
  • 抗原 See all G-CSF (CSF3) ELISA试剂盒
    G-CSF (CSF3) (Colony Stimulating Factor 3 (Granulocyte) (CSF3))
    适用
    • 8
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    • 1
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    检测方法
    Colorimetric
    实验类型
    Sandwich ELISA
    检测范围
    1-500 pg/mL
    最低检测浓度
    1 pg/mL
    应用范围
    ELISA
    原理
    Human GCSF ELISA Kit for cell culture supernatants, plasma, and serum samples.
    样品类型
    Plasma, Cell Culture Supernatant, Serum
    Analytical Method
    Quantitative
    特异性
    This ELISA kit shows no cross-reactivity with the following cytokines tested: human Angiogenin, 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, GM-CSF, IFN-gamma, Leptin, 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.
    灵敏度
    < 1 pg/mL
    产品特性
    • 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
    组件
    • 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
    试剂未包括
    • 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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  • 应用备注
    Recommended Dilution for serum and plasma samples2 fold
    样本量
    100 μL
    板类型
    Pre-coated
    实验流程
    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.
    试剂准备
    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, and Assay Diluent B (Item E) should be used for dilution of culture supernatants and urine. Suggested dilution for normal serum/plasma: 2 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 800 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 10 ng/mL standard. Dissolve the powder thoroughly by a gentle mix. Add 30 µL G-CSF standard from the vial of Item C, into a tube with 570 µL Assay Diluent A or 1x Assay Diluent B to prepare a 500 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 200 µL 200 µL 200 µL 200 µL 30 µL standard + 570 µL 200myl 500 166.7 55.56 18.52 6.17 2.05 0.69 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 260-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 13 ml 1x Assay Diluent B to prepare a final 260 fold diluted HRP-Streptavidin solution (don't store the diluted solution for next day use). Mix well.
    实验流程
    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.
    结果分析

    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. Human G-CSF concentration (pg/mL) 0.1 1 10 100 1000 O D =4 50 n m 0.01 0.1 1 10 Assay Diluent A Human G-CSF concentration (pg/mL) 0.1 1 10 100 1000 O D =4 50 n m 0.01 0.1 1 10 Assay Diluent B
    Sensitivity: The minimum detectable dose of G-CSF is typically less than 1 pg/mL.
    Recovery: Recovery was determined by spiking human G-CSF into human serum, plasma and cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Serum 83.45 72-94 Plasma 74.76 67-87 Cell culture media 76.56 68-90
    Linearity: Sample Type Serum Plasma Cell Culture Media 1:2 Average % of Expected 128.8 134.7 109.3 Range ( %) 117-136 120-139 98-117 1:4 Average % of Expected 134.3 137.8 108.1 Range ( %) 123-139 124-143 97-115
    Reproducibility: Intra-Assay: CV<10 % Inter-Assay: CV<12 %

    实验精密度
    Intra-Assay: CV< 10 % Inter-Assay: CV< 12 %
    限制
    仅限研究用
  • 注意事项
    Avoid repeated freeze-thaw cycles.
    储存条件
    -20 °C
    储存方法
    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.
    有效期
    6 months
  • Lee, Shynlova, Lye: "Stretch-induced human myometrial cytokines enhance immune cell recruitment via endothelial activation." in: Cellular & molecular immunology, Vol. 12, Issue 2, pp. 231-42, (2015) (PubMed).

    Koob, Lim, Zabek, Massee: "Cytokines in single layer amnion allografts compared to multilayer amnion/chorion allografts for wound healing." in: Journal of biomedical materials research. Part B, Applied biomaterials, Vol. 103, Issue 5, pp. 1133-40, (2015) (PubMed).

    Wu, Liu, Huang, Chen, Chang, Ho, Chao: "A phase II double-blinded study to evaluate the efficacy of EW02 in reducing chemotherapy-induced neutropenia in breast cancer." in: Oncology letters, Vol. 10, Issue 3, pp. 1793-1798, (2015) (PubMed).

    Koob, Lim, Massee, Zabek, Denozière: "Properties of dehydrated human amnion/chorion composite grafts: Implications for wound repair and soft tissue regeneration." in: Journal of biomedical materials research. Part B, Applied biomaterials, Vol. 102, Issue 6, pp. 1353-62, (2014) (PubMed).

    Nair, Chidambareswaren, Manjula: "Enhanced heterologous expression of biologically active human granulocyte colony stimulating factor in transgenic tobacco BY-2 cells by localization to endoplasmic reticulum." in: Molecular biotechnology, Vol. 56, Issue 9, pp. 849-62, (2014) (PubMed).

    Batista, Melo, Souza-Fabjan, Teixeira, Melo, Freitas: "Phenotypic features of first-generation transgenic goats for human granulocyte-colony stimulation factor production in milk." in: Biotechnology letters, Vol. 36, Issue 11, pp. 2155-62, (2014) (PubMed).

    El-Ganzoury, Awad, El-Farrash, El-Gammasy, Ismail, Mohamed, Suliman: "Enteral granulocyte-colony stimulating factor and erythropoietin early in life improves feeding tolerance in preterm infants: a randomized controlled trial." in: The Journal of pediatrics, Vol. 165, Issue 6, pp. 1140-1145.e1, (2014) (PubMed).

    Moura, Albuquerque, Melo, Alcântara-Neto, Batista, Nunes-Pinheiro, Pereira, Teixeira, Melo, Serova, Andreeva, Serov, Freitas: "Dynamics of recombinant hG-CSF in transgenic goat: preliminary study in the founder during hormonally induced lactation." in: Animal biotechnology, Vol. 24, Issue 1, pp. 10-4, (2013) (PubMed).

    Alrokayan: "Chemical synthesis and improved expression of recombinant human granulocyte colony-stimulating factor cDNA." in: Genetics and molecular research : GMR, Vol. 10, Issue 4, pp. 2671-8, (2012) (PubMed).

    Shirasaki, Tashiro, Mizutani-Noguchi, Kawasugi, Shirafuji: "Chronic myelogenous leukemia cells convert to myofibroblasts in vitro: effect of vascular endothelial growth factor on development of the microenvironment." in: Leukemia research, Vol. 35, Issue 5, pp. 663-9, (2011) (PubMed).

    Harada, Ohuchi, Hayashi, Kato: "Prolonged circulation and in vivo efficacy of recombinant human granulocyte colony-stimulating factor encapsulated in polymeric micelles." in: Journal of controlled release : official journal of the Controlled Release Society, Vol. 156, Issue 1, pp. 101-8, (2011) (PubMed).

    Maekawa, Sonoda, Kuzuyama, Inazawa, Kimura, Nakamichi, Abe: "Synergistic suppression of the clonogenicity of U937 leukemic cells by combinations of recombinant human interleukin 4 and granulocyte colony-stimulating factor." in: Experimental hematology, Vol. 20, Issue 10, pp. 1201-7, (1992) (PubMed).

    Moore: "The clinical use of colony stimulating factors." in: Annual review of immunology, Vol. 9, pp. 159-91, (1991) (PubMed).

  • 抗原 See all G-CSF (CSF3) ELISA试剂盒
    G-CSF (CSF3) (Colony Stimulating Factor 3 (Granulocyte) (CSF3))
    别名
    G-CSF / CSF3 (CSF3 产品)
    别名
    CSF3 ELISA Kit, G-CSF ELISA Kit, gcsf ELISA Kit, Csfg ELISA Kit, MGI-IG ELISA Kit, C17orf33 ELISA Kit, CSF3OS ELISA Kit, GCSF ELISA Kit, Gcsf ELISA Kit, colony stimulating factor 3 ELISA Kit, colony stimulating factor 3 (granulocyte) a ELISA Kit, colony stimulating factor 3 (granulocyte) ELISA Kit, CSF3 ELISA Kit, csf3a ELISA Kit, Csf3 ELISA Kit
    背景
    G-CSF (Granulocyte colony stimulating factor), which is an O-glycosylated 19.6 kDa glycoprotein, is secreted by monocytes, macrophages and neutrophils after cell activation. G-CSF is produced also by stromal cells, fibroblasts, and endothelial cells. It stimulates the proliferation and differentiation of hematopoietic progenitor cells committed to the neutrophil/granulocyte lineage in a dose-dependent manner. A combination of IL-4 with G-CSF has been shown to lead to synergistic suppression of the growth of some human leukemic cell lines. The main and most important clinical application of G-CSF is probably the treatment of transient phases of leukopenia following chemotherapy and/or radiotherapy. The Human G-CSF ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of human G-CSF in serum, plasma, cell culture supernatants and urine. This assay employs an antibody specific for human G-CSF coated on a 96-well plate. Standards and samples are pipetted into the wells and G-CSF present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-human G-CSF 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 G-CSF 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%.
    基因ID
    1440
    UniProt
    P09919
    途径
    Cellular Response to Molecule of Bacterial Origin, Regulation of Actin Filament Polymerization
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