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BDNF ELISA Kit

BDNF Reactivity: Human Colorimetric Sandwich ELISA 80 pg/mL-16 ng/mL Cell Culture Supernatant, Plasma, Serum
Catalog No. ABIN624945
  • Target See all BDNF ELISA Kits
    BDNF (Brain-Derived Neurotrophic Factor (BDNF))
    Reactivity
    • 9
    • 6
    • 5
    • 4
    • 3
    • 3
    • 3
    • 3
    • 2
    • 2
    • 2
    • 2
    • 2
    • 1
    Human
    Detection Method
    Colorimetric
    Method Type
    Sandwich ELISA
    Detection Range
    80 pg/mL-16 ng/mL
    Minimum Detection Limit
    80 pg/mL
    Application
    ELISA
    Purpose
    Human BDNF 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, 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-1delta, PARC, PDGF, RANTES, SCF, TARC, TGF- beta, TIMP-1, TIMP-2, TNF- alpha, TNF- beta, TPO, VEGF.
    Sensitivity
    80 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 samples5 - 500 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: 10-500 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 720 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 400 ng/mL standard. Dissolve the powder thoroughly by a gentle mix. Add 40 µL BDNF standard from the vial of Item C, into a tube with 960 µL Assay Diluent A or 1x Assay Diluent B to prepare a 16 ng/mL 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 ng/mL). 200 µL 200myl 200 µL 200 µL 200 µL 200 µL 40 µL standard + 960 µL 16 6.4 2.56 1.02 0.41 0.16 0.066 0 ng/mL ng/mL ng/mL ng/mL ng/mL ng/mL ng/mL ng/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 200-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 10 ml 1x Assay Diluent B to prepare a final 200 fold diluted HRP- Streptavidin solution (don't store the diluted solution for next day use). 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 BDNF concentration (ng/mL) 0.01 0.1 1 10 100 O D =4 50 (n m ) 0.1 1 10 Assay Diluent B Human BDNF concentration (ng/mL) 0.01 0.1 1 10 100 O D =4 50 (n m ) 0.01 0.1 1 10
    Sensitivity: The minimum detectable dose of BDNF is typically less than 80 pg/mL.
    Recovery: Recovery was determined by spiking various levels of human BDNF into human serum, plasma and cell culture media. Mean recoveries are as follows: Sample Type Average % Recovery Range ( %) Serum 94.57 83-103 Plasma 92.48 82-102 Cell culture media 93.35 83-103
    Linearity: Sample Type Serum Plasma Cell Culture Media 1:2 Average % of Expected 93 92 94 Range ( %) 83-103 84-103 83-104 1:4 Average % of Expected 94 93 95 Range ( %) 84-104 83-102 85-103
    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
  • van den Heuvel, Suliman, Malan-Müller, Hemmings, Seedat: "Brain-derived neurotrophic factor Val66met polymorphism and plasma levels in road traffic accident survivors." in: Anxiety, stress, and coping, Vol. 29, Issue 6, pp. 616-29, (2018) (PubMed).

    Cadenas-Sánchez, Mora-González, Migueles, Martín-Matillas, Gómez-Vida, Escolano-Margarit, Maldonado, Enriquez, Pastor-Villaescusa, de Teresa, Navarrete, Lozano, de Dios Beas-Jiménez, Estévez-López et al.: "An exercise-based randomized controlled trial on brain, cognition, physical health and mental health in overweight/obese children (ActiveBrains project): Rationale, design and methods. ..." in: Contemporary clinical trials, Vol. 47, pp. 315-24, (2017) (PubMed).

    Failla, Juengst, Arenth, Wagner: "Preliminary Associations Between Brain-Derived Neurotrophic Factor, Memory Impairment, Functional Cognition, and Depressive Symptoms Following Severe TBI." in: Neurorehabilitation and neural repair, Vol. 30, Issue 5, pp. 419-30, (2016) (PubMed).

    Faroni, Smith, Lu, Reid: "Human Schwann-like cells derived from adipose-derived mesenchymal stem cells rapidly de-differentiate in the absence of stimulating medium." in: The European journal of neuroscience, Vol. 43, Issue 3, pp. 417-30, (2016) (PubMed).

    Sadowska-Krępa, Zwierzchowska, Głowacz, Borowiec-Rybak, Kłapcińska: "Blood metabolic response to a long-term wheelchair rugby training." in: Spinal cord, Vol. 54, Issue 5, pp. 371-5, (2016) (PubMed).

    Wysokiński: "Serum levels of brain-derived neurotrophic factor (BDNF) and neurotrophin-3 (NT-3) in depressed patients with schizophrenia." in: Nordic journal of psychiatry, Vol. 70, Issue 4, pp. 267-71, (2016) (PubMed).

    Liu, Jiao, Wang, Bu, Yao, Xiang, Wang, Wang, Deng, Li, Zhou, Zhou, Wang: "Associations Between ApoE?4 Carrier Status and Serum BDNF Levels--New Insights into the Molecular Mechanism of ApoE?4 Actions in Alzheimer's Disease." in: Molecular neurobiology, Vol. 51, Issue 3, pp. 1271-7, (2015) (PubMed).

    Fornaro, Escelsior, Rocchi, Conio, Magioncalda, Marozzi, Presta, Sterlini, Contini, Amore, Fornaro, Martino: "BDNF plasma levels variations in major depressed patients receiving duloxetine." in: Neurological sciences : official journal of the Italian Neurological Society and of the Italian Society of Clinical Neurophysiology, Vol. 36, Issue 5, pp. 729-34, (2015) (PubMed).

    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).

    Lu, Zhang, Wen, Sun: "Impact of repetitive transcranial magnetic stimulation on post-stroke dysmnesia and the role of BDNF Val66Met SNP." in: Medical science monitor : international medical journal of experimental and clinical research, Vol. 21, pp. 761-8, (2015) (PubMed).

    Oral, Kirkan, Yildirim, Kotan, Cansever, Ozcan, Aliyev, Gulec: "Serum brain-derived neurotrophic factor differences between the luteal and follicular phases in premenstrual dysphoric disorder." in: General hospital psychiatry, Vol. 37, Issue 3, pp. 266-72, (2015) (PubMed).

    Jiang, Ling, Zhang, Mao, Ma, Yin, Wang, Tang, Tan, Shi, Li, Ruan: "Altered fecal microbiota composition in patients with major depressive disorder." in: Brain, behavior, and immunity, Vol. 48, pp. 186-94, (2015) (PubMed).

    Failla, Conley, Wagner: "Brain-Derived Neurotrophic Factor (BDNF) in Traumatic Brain Injury-Related Mortality: Interrelationships Between Genetics and Acute Systemic and Central Nervous System BDNF Profiles." in: Neurorehabilitation and neural repair, Vol. 30, Issue 1, pp. 83-93, (2015) (PubMed).

    Haslacher, Michlmayr, Batmyagmar, Perkmann, Ponocny-Seliger, Scheichenberger, Pilger, Dal-Bianco, Lehrner, Pezawas, Wagner, Winker: "Physical exercise counteracts genetic susceptibility to depression." in: Neuropsychobiology, Vol. 71, Issue 3, pp. 168-75, (2015) (PubMed).

    Eleftheriou, Ganesan, Hong, Klein, Brogan: "Endothelial Repair in Childhood Arterial Ischaemic Stroke with Cerebral Arteriopathy." in: Cerebrovascular diseases extra, Vol. 5, Issue 2, pp. 68-74, (2015) (PubMed).

    Zhang, Lin, Jiang, Xu, Luo, Mo, Li, Chen: "Extensive serum biomarker analysis in patients with ST segment elevation myocardial infarction (STEMI)." in: Cytokine, Vol. 76, Issue 2, pp. 356-62, (2015) (PubMed).

    Kielstein, Suntharalingam, Perthel, Song, Schneider, Martens-Lobenhoffer, Jäger, Bode-Böger, Kielstein et al.: "Role of the endogenous nitric oxide inhibitor asymmetric dimethylarginine (ADMA) and brain-derived neurotrophic factor (BDNF) in depression and behavioural changes: clinical and preclinical data in ..." in: Nephrology, dialysis, transplantation : official publication of the European Dialysis and Transplant Association - European Renal Association, Vol. 30, Issue 10, pp. 1699-705, (2015) (PubMed).

    Schendzielorz, Rak, Nguyen, Frölich, Scherzad, Hagen, Radeloff: "Human adipose-derived stem cells enhance the survival and neuritogenesis of auditory neurons." in: Neuroreport, Vol. 26, Issue 13, pp. 797-801, (2015) (PubMed).

    Li, He, Wu, Pan, Wang, Tan, Shan, Zeng: "PFOS Disturbs BDNF-ERK-CREB Signalling in Association with Increased MicroRNA-22 in SH-SY5Y Cells." in: BioMed research international, Vol. 2015, pp. 302653, (2015) (PubMed).

    Sahin, Yuce, Alacam, Karabekiroglu, Say, Sal?s et al.: "Effect of methylphenidate treatment on appetite and levels of leptin, ghrelin, adiponectin, and brain-derived neurotrophic factor in children and adolescents with attention deficit and hyperactivity ..." in: International journal of psychiatry in clinical practice, Vol. 18, Issue 4, pp. 280-7, (2014) (PubMed).

  • Target See all BDNF ELISA Kits
    BDNF (Brain-Derived Neurotrophic Factor (BDNF))
    Alternative Name
    BDNF (BDNF Products)
    Synonyms
    ANON2 ELISA Kit, BULN2 ELISA Kit, bdnf-A ELISA Kit, BDNF ELISA Kit, bdnf ELISA Kit, brain derived neurotrophic factor ELISA Kit, brain-derived neurotrophic factor ELISA Kit, brain-derived neurotrophic factor L homeolog ELISA Kit, brain-derived neurotrophic factor S homeolog ELISA Kit, BDNF ELISA Kit, Bdnf ELISA Kit, bdnf ELISA Kit, bdnf.L ELISA Kit, bdnf.S ELISA Kit
    Background
    BDNF is found in neurons of the central nervous system. It is expressed predominantly in hippocampus, cortex, and synapses of the basal forebrain. BDNF selectively supports the survival of primary sensory neurons and retinal ganglia. It supports survival and differentiation of certain cholinergic neurons and also some dopaminergic neurons in vitro. The factor inhibits the normal cell death of embryonic chick motor neurons. The Human BDNF ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of human BDNF in serum, plasma, cell culture supernatants and urine. This assay employs an antibody specific for human BDNF coated on a 96-well plate. Standards and samples are pipetted into the wells and BDNF present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-human BDNF 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 BDNF 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
    627
    UniProt
    P23560
    Pathways
    RTK Signaling, Synaptic Membrane, Feeding Behaviour, Dicarboxylic Acid Transport, Regulation of long-term Neuronal Synaptic Plasticity
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