Total BDNF ELISA Kit - Quantikine Best Seller
R&D Systems, part of Bio-Techne | Catalog # DBNT00
Detects free and Trk-bound BDNF
Key Product Details
Assay Length
3.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 µL), Tissue Lysates (50 µL), Serum (10 µL), EDTA Plasma (50 µL), Platelet-poor EDTA Plasma (10 µL), Heparin Plasma (50 µL), Platelet-poor Heparin Plasma (10 µL), Urine (50 µL), Human Milk (50 µL)
Sensitivity
1.35 pg/mL
Assay Range
15.6-1000 pg/mL (Cell Culture Supernates, Tissue Lysates, Serum, EDTA Plasma, Platelet-poor EDTA Plasma, Heparin Plasma, Platelet-poor Heparin Plasma, Urine, Human Milk)
Product Summary for Total BDNF Quantikine ELISA Kit
The Quantikine Total BDNF Immunoassay is a 3.5 hour solid phase ELISA designed to measure BDNF in cell culture supernates, tissue lysates, serum, plasma, urine, and human milk. 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 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 BDNF.
Product Specifications
Measurement
Quantitative ELISA
Detection Method
Colorimetric - 450nm (TMB)
Conjugate
HRP
Reactivity
Human, Mouse, Rat, Porcine, Canine
Specificity
Natural and recombinant free, trk-bound, pro-, and mature BDNF.
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules. < 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.
Sample Values
Serum/Plasma/Urine/Human milk - Samples from apparently healthy volunteers were
evaluated for the presence of Total BDNF in this assay. No medical histories were available for
the donors used in this study.
ND=Non-detectable
Human peripheral blood mononuclear cells (50 x 106 cells/mL) were cultured in RPMI supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate overnight. The cells were cultured unstimulated or stimulated with 1.0 μg/mL LPS for 1 day. Aliquots of the cell culture supernates were removed, assayed for levels of Total BDNF, and measured 2870 pg/mL and 2853 pg/mL, respectively.
Tissue lysates:
Brains from mice were rinsed with PBS and homogenized with a tissue homogenizer in PBS. An equal volume of Lysis Buffer 17 was added and tissues were lysed at room temperature for 30 minutes with gentle agitation. Debris was then removed by centrifugation. An aliquot of each cell lysate was removed, assayed for levels of Total BDNF, and measured 1472 pg/mg of cell lysate.
| Sample Type | Mean (pg/mL) | Range (pg/mL) | Standard Deviation (pg/mL) |
| Human serum (n=36) | 35,615 | 9042-57,830 | 9790 |
| Human PP EDTA plasma (n=36) | 307 | 94.9-902 | 197 |
| Human PP heparin plasma (n=36) | 595 | 100-2245 | 491 |
| Rat serum (n=10) | 5253 | 2554-6966 | 1438 |
| Rat PP EDTA plasma (n=5) | 294 | 87.5-416 | 138 |
| Rat PP heparin plasma (n=5) | 294 | 103-713 | 248 |
| Sample Type | Mean of Detectable (pg/mL) | % Detectable | Range (pg/mL) |
| Human Urine (n=20) | ND | 0 | ND |
| Human milk (n=10) | ND | 0 | ND |
| Mouse serum (n=10) | 55.4 | 50 | ND-147 |
| Mouse PP EDTA plasma (n=5) | 21.4 | 60 | ND-29.7 |
| Mouse PP heparin plasma (n=5) | 17.5 | 40 | ND-19.1 |
| Mouse urine (n=5) | ND | 0 | ND |
| Rat urine (n=5) | ND | 0 | ND |
| Canine serum (n=4) | ND | 0 | ND |
| Canine EDTA plasma (n=4) | ND | 0 | ND |
| Canine heparin plasma (n=4) | ND | 0 | ND |
| Porcine serum (n=4) | ND | 0 | ND |
| Porcine EDTA plasma (n=4) | ND | 0 | ND |
| Porcine heparin plasma (n=4) | ND | 0 | ND |
Cell Culture Supernates:
Human peripheral blood leukocytes (1 x 106 cells) were cultured in RPMI supplemented with
10% fetal bovine serum. The cells were cultured unstimulated or stimulated with 10 μg/mL
PHA for 1 and 5 days. Aliquots of the cell culture supernates were removed and assayed for
levels of Total BDNF.
| Condition | Day 1 (pg/mL) | Day 5 (pg/mL) |
| Unstimulated | 25.5 | 21.2 |
| Stimulated | 24.4 | 24.4 |
Human peripheral blood mononuclear cells (50 x 106 cells/mL) were cultured in RPMI supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate overnight. The cells were cultured unstimulated or stimulated with 1.0 μg/mL LPS for 1 day. Aliquots of the cell culture supernates were removed, assayed for levels of Total BDNF, and measured 2870 pg/mL and 2853 pg/mL, respectively.
MG-63 human osteosarcoma cells were cultured in DMEM supplemented with 5% fetal bovine
serum and 4 mM L-glutamine and grown until confluent. An aliquot of the cell culture
supernate was removed, assayed for Total BDNF, and measured 156 pg/mL.
Brain tissues from mice and rats were removed and rinsed in PBS and kept on ice. The tissue
was homogenized using a tissue homogenizer and seeded into media containing RPMI 1640
supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 100 U/mL penicillin, and
100 μg/mL streptomycin sulfate. Cells were cultured unstimulated for 2 or 3 days. Aliquots of
the cell culture supernates were removed and assayed for levels of Total BDNF.
| Sample Type | Observed Levels (pg/mL) |
| Mouse brain (2 days) | 306 |
| Rat brain (3 days) | 196 |
Tissue lysates:
Human brain tissues were prepared in RIPA buffer with protease inhibitors. The working buffer
was kept on ice and used within 2-3 days. Aliquots of the tissue lysates were removed and
assayed for Total BDNF.
| Sample Type | pg/mg of Cell Lysate |
| Human cerebellum | 515 |
| Human hypothalamus | 441 |
Brains from mice were rinsed with PBS and homogenized with a tissue homogenizer in PBS. An equal volume of Lysis Buffer 17 was added and tissues were lysed at room temperature for 30 minutes with gentle agitation. Debris was then removed by centrifugation. An aliquot of each cell lysate was removed, assayed for levels of Total BDNF, and measured 1472 pg/mg of cell lysate.
Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components.
Cell Culture Supernates, EDTA Plasma, Heparin Plasma, Human Milk, Platelet-poor EDTA Plasma, Platelet-poor Heparin Plasma, Serum, Tissue Lysates, Urine
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 97.9 | 311 | 586 | 98.8 | 296 | 594 |
| Standard Deviation | 3.12 | 7.52 | 17.5 | 7.07 | 12.7 | 28.0 |
| CV% | 3.2 | 2.4 | 3.0 | 7.2 | 4.3 | 4.7 |
Recovery for Total BDNF Quantikine ELISA Kit
The recovery of Total BDNF spiked to levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Media (n=4) | 103 | 97-113 |
| Cell Lysis Buffer (n=4) | 94 | 87-100 |
| Human Milk (n=4) | 94 | 87-99 |
| Platelet-poor EDTA Plasma (n=4) | 95 | 76-110 |
| Platelet-poor Heparin Plasma (n=4) | 95 | 75-111 |
| Urine (n=4) | 93 | 75-106 |
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of Total BDNF were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Scientific Data Images for Total BDNF Quantikine ELISA Kit
Human/Mouse/Rat/Porcine/Canine Total BDNF ELISA 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
Long Name
Brain-derived Neurotrophic Factor
Alternate Names
Abrineurin, ANON2, BULN2, Neurotrophin
Gene Symbol
BDNF
Additional BDNF Products
Product Documents for Total BDNF Quantikine ELISA Kit
Product Specific Notices for Total BDNF Quantikine ELISA Kit
For research use only
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⚠ 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.