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Human Free BDNF Quantikine ELISA Kit

R&D Systems | Catalog # DBD00

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Key Product Details

Assay Length

3.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (50 µL), Serum (10 µL), EDTA Plasma (50 µL), Platelet-poor Heparin Plasma (50 µL), Citrate Plasma (50 µL)

Sensitivity

20 pg/mL

Assay Range

62.5-4000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Platelet-poor Heparin Plasma, Citrate Plasma)
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Product Summary for Human Free BDNF Quantikine ELISA Kit

The Quantikine Human BDNF Immunoassay is a 3.5 hour solid phase ELISA designed to measure human BDNF in cell culture supernates, serum, and plasma. It contains recombinant human BDNF expressed in Sf 21 cells and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant BDNF accurately. Results obtained measuring natural human BDNF showed dose-response curves that were parallel to the standard curves obtained using the recombinant Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for natural human BDNF.

Product Specifications

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Species

Human

Specificity

Natural and recombinant human BDNF

Cross-reactivity

Cross-reactivity observed with 1 or more available related molecules. < 50% cross-species reactivity observed with species tested.

Interference

Interference observed with 1 or more available related molecules.

Sample Values

Serum/Plasma - Samples from apparently healthy volunteers were evaluated for the presence of human free BDNF in this assay. No medical histories were available for the donors used in this study.

Sample TypeMean of Detectable (pg/mL)% DetectableRange (pg/mL)
Serum* (n=33)27,7931006186-42,580
Platelet-poor EDTA plasma (n=33)42856ND-4137
Platelet-poor heparin plasma (n=33)19655ND-528
Platelet-poor citrate plasma (n=33)40194ND-1838
*Samples were diluted prior to assay.
 ND=Non-detectable

Cell Culture Supernates - Human peripheral blood  mononuclear cells (1 x 106 cells/mL) were cultured in RPMI supplemented with 10% fetal bovine serum, 50 μM beta -mercaptoethanol, 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate and stimulated with 10 μg/mL PHA. Aliquots of the cell culture supernates were removed on days 1 and 5 and assayed for levels of human free BDNF.

ConditionDay 1 (pg/mL)Day 5 (pg/mL)
Unstimulated132109
Stimulated160104



精度

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.

Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.

Cell Culture Supernates

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (pg/mL) 334 825 1339 358 905 1517
Standard Deviation 15.6 28.3 56.5 36.8 86.8 121.6
CV% 4.7 3.4 4.2 10.3 9.6 8.1

Citrate Plasma, EDTA Plasma, Platelet-poor Heparin Plasma, Serum

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean (pg/mL) 476 1258 2175 528 1338 2287
Standard Deviation 24.0 48.2 135.7 59.7 102.3 186.0
CV% 5.0 3.8 6.2 11.3 7.6 8.1

Recovery for Human Free BDNF Quantikine ELISA Kit

The recovery of BDNF spiked to three different levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=5) 96 83-105
Platelet-poor Citrate Plasma (n=5) 99 87-120
Platelet-poor EDTA Plasma (n=5) 96 89-115
Platelet-poor Heparin Plasma (n=5) 93 81-119
Serum (n=5) 100 85-116

Linearity

To assess the linearity of the assay, five samples containing or spiked with high concentrations of BDNF in various matrices were diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.

Human Free BDNF ELISA Linearity

Scientific Data Images for Human Free BDNF Quantikine ELISA Kit

Human Free BDNF ELISA Cell Culture Supernate Standard Curve

Human Free BDNF ELISA Cell Culture Supernate Standard Curve

Human Free BDNF ELISA Serum/Plasma Standard Curve

Human Free BDNF ELISA Serum/Plasma Standard Curve

Preparation and Storage

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: BDNF

BDNF (Brain-Derived Neurotrophic Factor) is a secreted protein that regulates many aspects of neuronal development and function in the nervous system. It is produced as proBDNF in response to neuronal activity or inflammatory stimulation and is then cleaved before associating into a homodimer. It is expressed throughout the nervous system as well as by fibroblasts, megakaryocytes/platelets, and smooth muscle cells. BDNF regulates neural stem cell survival and differentiation, axon/dendrite differentiation, synapse formation and maturation, and refinement of developing circuits. It is also a key regulator of synaptic plasticity and late-phase long-term potentiation. BDNF signals through the TrkB and NGF R/TNFRSF16 receptors. Secreted proBDNF can bind to NGF R/TNFRSF16 and induce long-term depression and cell apoptosis.

Long Name

Brain-derived Neurotrophic Factor

Alternate Names

Abrineurin, ANON2, BULN2, Neurotrophin

Entrez Gene IDs

627 (Human); 12064 (Mouse); 24225 (Rat)

Gene Symbol

BDNF

Additional BDNF Products

Product Documents for Human Free BDNF Quantikine ELISA Kit

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human Free BDNF Quantikine ELISA Kit

For research use only

⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

Citations for Human Free BDNF Quantikine ELISA Kit

Customer Reviews for Human Free BDNF Quantikine ELISA Kit (6)

4.7 out of 5
6 Customer Ratings
5 Stars
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4 Stars
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Showing  1 - 5 的 6 reviews Showing All
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  • Human Free BDNF Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Cell culture supernatant
    Verified Customer | Posted 11/15/2023
    Human Free BDNF Quantikine ELISA Kit DBD00
  • Human Free BDNF Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Serum and Plasma
    Verified Customer | Posted 11/12/2020
    Very good standard curve, samplesl worked at a dilution of 1:200
    Human Free BDNF Quantikine ELISA Kit DBD00
  • Human Free BDNF Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: EDTA Plasma
    Verified Customer | Posted 10/24/2020
    Human Free BDNF Quantikine ELISA Kit DBD00
  • Name: Ruby Argumedo
    Sample Tested: Heparin Plasma
    Verified Customer | Posted 12/05/2018
    We use this kit really frequently so a 6 pack of BDNF is very convenient for us to have. Easy to use and to understand. Results are usually really good and reproducible. Would definitely recommend.
  • Human Free BDNF Quantikine ELISA Kit
    Name: Anonymous
    Sample Tested: Serum
    Verified Customer | Posted 11/02/2018
  • Human Free BDNF Quantikine ELISA Kit
    Name: Luiz Alexandre
    Sample Tested: Heparin Plasma, EDTA Plasma and Mouse lymphocytes
    Verified Customer | Posted 07/13/2018
    Human Free BDNF Quantikine ELISA Kit DBD00

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Showing  1 - 5 的 6 reviews Showing All

Protocols

View specific protocols for Human Free BDNF Quantikine ELISA Kit (DBD00):

Refer to the product for complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
    3. 100 µL Assay Diluent
  3.   Add 100 µL of Assay Diluent to each well.
    4. 50 µL Standard, Control, or Sample
  4.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
    Go to the next step. Do not wash.
    5. 100 µL Conjugate
  5.   Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 1 hour.
  6.   Aspirate each well and wash, repeating the process twice for a total of 3 washes.
    7. 200 µL Substrate Solution
  7.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
    8. 50 µL Stop Solution
  8.   Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

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