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FGF2 ELISA Kit

FGF2 Reactivity: Human Colorimetric Sandwich ELISA 50-10000 pg/mL Cell Culture Supernatant, Plasma, Serum
Catalog No. ABIN624947
  • Target See all FGF2 ELISA Kits
    FGF2 (Fibroblast Growth Factor 2 (Basic) (FGF2))
    Reactivity
    • 10
    • 5
    • 3
    • 3
    • 3
    • 3
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    Human
    Detection Method
    Colorimetric
    Method Type
    Sandwich ELISA
    Detection Range
    50-10000 pg/mL
    Minimum Detection Limit
    50 pg/mL
    Application
    ELISA
    Purpose
    Human bFGF ELISA Kit for cell culture supernatants, plasma, and serum samples.
    Sample Type
    Plasma, Cell Culture Supernatant, Serum
    Analytical Method
    Quantitative
    Specificity
    This ELISA kit shows no cross-reactivity with any of the 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, G-CSF, 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.
    Sensitivity
    < 50 pg/mL
    Characteristics
    • 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
    Components
    • 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
    Material not included
    • 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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  • Application Notes
    Recommended Dilution for serum and plasma samples2 fold
    Sample Volume
    100 μL
    Plate
    Pre-coated
    Protocol
    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.
    Reagent Preparation
    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 supernantants 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 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 100 ng/mL standard. Dissolve the powder thoroughly by a gentle mix. Add 100 µL bFGF standard from the vial of Item C, into a tube with 900 µL Assay Diluent A or 1x Assay Diluent B to prepare a 10,000 pg/mL stock standard solution. Pipette 300 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 100 µL standard + 900 µL 200myl 200 µL 200 µL 200 µL 10,000 4,000 1,600 640 256 102.4 0 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 65-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 120-fold with 1x Assay Diluent B. 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 1x Assay Diluent B tocprepare a final 120 fold diluted HRP-Streptavidin solution. Mix well.
    Assay Procedure
    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.
    Calculation of Results

    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 bFGF concentration (pg/mL) O D =4 50 n m 0.001 0.01 0.1 1 10 10 100 1,000 10,000 Assay Diluent B Human bFGF concentration (pg/mL) O D =4 50 n m 0.001 0.01 0.1 1 10 10 100 1,000 10,000
    Sensitivity: The minimum detectable dose of bFGF is typically less than 50 pg/mL.
    Recovery: Recovery was determined by spiking various levels of human bFGF into human serum, plasma and cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Serum 95.46 84-104 Plasma 92.64 82-103 Cell culture media 93.48 83-103
    Linearity: Sample Type Serum Plasma Cell Culture Media 1:2 Average % of Expected 91 89 92 Range ( %) 85-103 84-103 84-103 1:4 Average % of Expected 93 92 90 Range ( %) 83-104 83-104 84-105
    Reproducibility: Intra-Assay: CV<10 % Inter-Assay: CV<12 %

    Assay Precision
    Intra-Assay: CV< 10 % Inter-Assay: CV< 12 %
    Restrictions
    For Research Use only
  • Handling Advice
    Avoid repeated freeze-thaw cycles.
    Storage
    -20 °C
    Storage Comment
    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.
    Expiry Date
    6 months
  • Yamazoe, Hagihara, Kobayashi: "Multicomponent Coculture System of Cancer Cells and Two Types of Stromal Cells for In Vitro Evaluation of Anticancer Drugs." in: Tissue engineering. Part C, Methods, Vol. 22, Issue 1, pp. 20-9, (2016) (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).

    Guan, Guo, Yu, Wang, Wang, Konstantopoulos, Wang, Wang: "The role of cyclooxygenase-2, interleukin-1β and fibroblast growth factor-2 in the activation of matrix metalloproteinase-1 in sheared-chondrocytes and articular cartilage." in: Scientific reports, Vol. 5, pp. 10412, (2015) (PubMed).

    Sowmya, Chennazhi, Arzate, Jayachandran, Nair, Jayakumar: "Periodontal Specific Differentiation of Dental Follicle Stem Cells into Osteoblast, Fibroblast, and Cementoblast." in: Tissue engineering. Part C, Methods, (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).

    Koob, Lim, Massee, Zabek, Rennert, Gurtner, Li: "Angiogenic properties of dehydrated human amnion/chorion allografts: therapeutic potential for soft tissue repair and regeneration." in: Vascular cell, Vol. 6, pp. 10, (2014) (PubMed).

    Lai, Kuan, Wu, Tsai, Chen, Hsieh, Wang: "Tailored design of electrospun composite nanofibers with staged release of multiple angiogenic growth factors for chronic wound healing." in: Acta biomaterialia, Vol. 10, Issue 10, pp. 4156-66, (2014) (PubMed).

    Esmaeelinejad, Bayat: "Effect of low-level laser therapy on the release of interleukin-6 and basic fibroblast growth factor from cultured human skin fibroblasts in normal and high glucose mediums." in: Journal of cosmetic and laser therapy : official publication of the European Society for Laser Dermatology, Vol. 15, Issue 6, pp. 310-7, (2014) (PubMed).

    Bhaskar, Gupta, Vishnubhatla, Kumar, Sharma, Sharma, Das, Thakur: "Angiopoietins as biomarker of disease activity and response to therapy in multiple myeloma." in: Leukemia & lymphoma, Vol. 54, Issue 7, pp. 1473-8, (2014) (PubMed).

    Yalç?nda?, Gedik-O?uz, Yalç?nda?: "The relationship between serum levels of angiogenin, bFGF, VEGF, and ocular involvement in patients with Behçet's disease." in: Graefe's archive for clinical and experimental ophthalmology, Vol. 251, Issue 7, pp. 1807-12, (2013) (PubMed).

    Bhaskar, Gupta, Sreenivas, Rani, Kumar, Sharma, Sharma, Sharma, Thakur: "Synergistic effect of vascular endothelial growth factor and angiopoietin-2 on progression free survival in multiple myeloma." in: Leukemia research, Vol. 37, Issue 4, pp. 410-5, (2013) (PubMed).

    Chang, Hwang, Brafman, Hagan, Phung, Varghese: "Engineering cell-material interfaces for long-term expansion of human pluripotent stem cells." in: Biomaterials, Vol. 34, Issue 4, pp. 912-21, (2013) (PubMed).

    Gok, Erdem, Gogus, Yilmaz, Karadag, Simsek, Sagkan, Saglam, Musabak, Dinc, Pay: "Relationship of ultrasonographic findings with synovial angiogenesis modulators in different forms of knee arthritides." in: Rheumatology international, Vol. 33, Issue 4, pp. 879-85, (2013) (PubMed).

    Fedak, Bai, Turnbull, Ngu, Narine, Duff: "Cell therapy limits myofibroblast differentiation and structural cardiac remodeling: basic fibroblast growth factor-mediated paracrine mechanism." in: Circulation. Heart failure, Vol. 5, Issue 3, pp. 349-56, (2012) (PubMed).

    Chim, Armijo, Miller, Gliniak, Serret, Gosain: "Propranolol induces regression of hemangioma cells through HIF-1?-mediated inhibition of VEGF-A." in: Annals of surgery, Vol. 256, Issue 1, pp. 146-56, (2012) (PubMed).

    Brown, Lau, Dom, Witte, Luo, Crabtree, Shah, Shiflett, Marcelo, Proper, Hardman, Egleton, Chen, Mangiarua, Dasgupta: "MG624, an ?7-nAChR antagonist, inhibits angiogenesis via the Egr-1/FGF2 pathway." in: Angiogenesis, Vol. 15, Issue 1, pp. 99-114, (2012) (PubMed).

    Nazarov, Lee, Soupene, Etemad, Knapik, Green, Bashkirova, Fang, Matthay, Kuypers, Serikov: "Multipotent stromal stem cells from human placenta demonstrate high therapeutic potential." in: Stem cells translational medicine, Vol. 1, Issue 5, pp. 359-72, (2012) (PubMed).

    Saygun, Nizam, Ural, Serdar, Avcu, Tözüm: "Low-level laser irradiation affects the release of basic fibroblast growth factor (bFGF), insulin-like growth factor-I (IGF-I), and receptor of IGF-I (IGFBP3) from osteoblasts." in: Photomedicine and laser surgery, Vol. 30, Issue 3, pp. 149-54, (2012) (PubMed).

    Kühn, Willenberg, Schott, Papewalis, Stumpf, Flohé, Scherbaum, Schinner: "Adipocyte-secreted factors increase osteoblast proliferation and the OPG/RANKL ratio to influence osteoclast formation." in: Molecular and cellular endocrinology, Vol. 349, Issue 2, pp. 180-8, (2011) (PubMed).

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  • Target See all FGF2 ELISA Kits
    FGF2 (Fibroblast Growth Factor 2 (Basic) (FGF2))
    Alternative Name
    bFGF (FGF2 Products)
    Synonyms
    BFGF ELISA Kit, FGF-2 ELISA Kit, FGFB ELISA Kit, HBGF-2 ELISA Kit, Fgf-2 ELISA Kit, Fgfb ELISA Kit, bFGF ELISA Kit, fibroblast growth factor 2 ELISA Kit, fibroblast growth factor 2 (basic) ELISA Kit, FGF2 ELISA Kit, Fgf2 ELISA Kit, fgf2 ELISA Kit
    Background
    BFGF (basic fibroblast growth factor) is found in almost all tissues of mesodermal and neuroectodermal origin and also in tumors derived from these tissues. Endothelial cells produce large amounts of this factor. Some bFGF is associated with the extracellular matrix of the subendothelial cells. bFGF is an 18 kDa protein with a length of 155 amino acids and an isoelectric point of 9.6. It does not contain disulfide bonds and is not glycosylated. bFGF stimulates the growth of fibroblasts, myoblasts, osteoblasts, neuronal cells, endothelial cells, keratinocytes, chondrocytes, and many other cell types. bFGF has been shown to be a promoting or inhibitory modulator of cellular differentiation also for other cell types. bFGF is not only a mitogen for chondrocytes but also inhibits their terminal differentiation. The Human bFGF ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of human bFGF in serum, plasma, cell culture supernatants and urine. This assay employs an antibody specific for human bFGF coated on a 96-well plate. Standards and samples are pipetted into the wells and bFGF present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-human bFGF 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 bFGF 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%.
    Gene ID
    2247
    UniProt
    P09038
    Pathways
    RTK Signaling, Fc-epsilon Receptor Signaling Pathway, EGFR Signaling Pathway, Neurotrophin Signaling Pathway, C21-Steroid Hormone Metabolic Process, Inositol Metabolic Process, Glycosaminoglycan Metabolic Process, Protein targeting to Nucleus, S100 Proteins
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