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Product Summary
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.
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 Type | Mean of Detectable (pg/mL) | % Detectable | Range (pg/mL) |
| Serum* (n=33) | 27,793 | 100 | 6186-42,580 |
| Platelet-poor EDTA plasma (n=33) | 428 | 56 | ND-4137 |
| Platelet-poor heparin plasma (n=33) | 196 | 55 | ND-528 |
| Platelet-poor citrate plasma (n=33) | 401 | 94 | ND-1838 |
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.
| Condition | Day 1 (pg/mL) | Day 5 (pg/mL) |
| Unstimulated | 132 | 109 |
| Stimulated | 160 | 104 |
Recovery
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.
Scientific Data
Assay Procedure
Refer to the product Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
- Prepare all reagents, standard dilutions, and samples as directed in the product insert.
- Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
- Add 100 µL of Assay Diluent to each well.
- 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. - Add 100 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 1 hour.
- Aspirate each well and wash, repeating the process twice for a total of 3 washes.
- Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
- 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.
100 µL Assay Diluent
50 µL Standard, Control, or Sample
100 µL Conjugate
200 µL Substrate Solution
50 µL Stop Solution
Human Free BDNF Quantikine ELISA Kit Summary
Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
3.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Serum (10 uL), EDTA Plasma (50 uL), Platelet-poor Heparin Plasma (50 uL), Citrate Plasma (50 uL)
Sensitivity
20 pg/mL
Assay Range
62.5 - 4,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Platelet-poor Heparin Plasma, Citrate Plasma)
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.
背景
别名
Abrineurin,ANON2,BDNF,brain-derived neurotrophic factor,BULN2,MGC34632,Neurotrophin,Brain-derived Neurotrophic Factor
背景
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
Entrez Gene IDs:
627 (Human); 12064 (Mouse); 24225 (Rat)
Alternate Names:
Abrineurin; ANON2; BDNF; brain-derived neurotrophic factor; BULN2; MGC34632; Neurotrophin
研究领域
制备和贮存
保存方式
Preparation and Storage
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
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