BDNF ELISA Kit

Catalog No. ABIN1112562

This Rat BDNF ELISA Kit is a Colorimetric ELISA Kit designed to quantify Rat BDNF.

Quick Overview for BDNF ELISA Kit (ABIN1112562)

Target: BDNF (Brain-Derived Neurotrophic Factor (BDNF))

Reactivity: Rat

Detection Method: Colorimetric

Method Type: Sandwich ELISA

Detection Range: 31.2-2000 pg/mL

Application: ELISA

Sample Type: Cell Culture Supernatant, Plasma, Serum, Tissue Lysate

Product Details BDNF ELISA Kit

Minimum Detection Limit: 31.2 pg/mL

Purpose: For quantitative detection of BDNF in Rat serum, plasma, body fluids, tissue lysates or cell culture supernatants.

Analytical Method: Quantitative

Sensitivity: < 2 pg/mL

Components: 1. One 96-well plate pre-coated with anti-Rat BDNF antibody 2. Lyophilized rat BDNF standards: 2 tubes (10ng / tube) 3. Sample / Standard diluent buffer: 30ml 4. Biotin conjugated anti-rat BDNF antibody (Concentrated): 130 µl.

Material not included: 1. 37 °C incubator 2. Microplate reader (wavelength: 450nm) 3. Precise pipette and disposable pipette tips 4. Automated plate washer 5. ELISA shaker 6. 1.5ml of Eppendorf tubes 7. Plate cover 8. Absorbent filter papers 9. Plastic or glass container with volume of above 1L

Application Details

Comment:

This kit was based on sandwich enzyme-linked immune-sorbent assay technology. Anti-BDNF polyclonal antibody was pre-coated onto 96-well plates. And the biotin conjugated anti-BDNF polyclonal antibody was used as detection antibodies. The standards test samples and biotin conjugated detection antibody were added - the wells subsequently and wash with wash buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with wash buffer. TMB substrates were used - visualize HRP enzymatic reaction. TMB was catalyzed by HRP - produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional - the BDNF amount of sample captured in plate. Read the O.D. absorbance at 450 nm in a microplate reader and then the concentration of BDNF can be calculated.

Plate: Pre-coated

Reagent Preparation:

1. Before the experiment, centrifuge each kit component for several minutes to bring down all reagents to the bottom of tubes. 2. It is recommend to measure each standard and sample in duplicate. 3. Do NOT let the plate completely dry at any time! Since the dry condition can inactivate the biological material on the plate. 4. Do not reuse pipette tips and tubes to avoid cross contamination. 5. Do not use the expired components and the components from different batches. 6. To avoid the marginal effect of plate incubation for temperature differences (the marginal wells always get stronger reaction), it is recommend to equilibrate the ABC working solution and TMB substrate for at least 30 min at room temperature (37°C ) before adding to wells.The TMB substrate (Kit Component 8) is colorless and transparent before use, if not, please contact us for replacement.

Sample Preparation:

Preparation of sample and reagents 1. Sample Isolate the test samples soon after collecting, then, analyze immediately (within 2 hours). Or aliquot and store at -20 °C for long term. Avoid multiple freeze-thaw cycles.
Body fluids, tissue lysates and cell culture supernatants: Centrifuge to remove precipitate, analyze immediately or aliquot and store at -20 °C .
Serum: Coagulate the serum at room temperature (about 4 hours). Centrifuge at approximately 1000 × g for 15 min. Analyze the serum immediately or aliquot and store at -20 °C .
Plasma: Collect plasma with citrate, heparin or EDTA as the anticoagulant. Centrifuge for 15min at 2-8°C at 1000 x g within 30 min of collection. For eliminating the platelet effect, suggesting that further centrifugation for 10 min at 2-8°C at 10000 x g. Analyze immediately or aliquot and store frozen at -20 °C. Note: 1. Coagulate blood samples completely, then, centrifuge, and avoid hemolysis and particle. 2. NaN3 can not be used as test sample preservative, since it is the inhibitor for HRP.

Restrictions: For Research Use only

Handling

Preservative: Sodium azide, Thimerosal (Merthiolate)

Target Details for BDNF

Target: BDNF (Brain-Derived Neurotrophic Factor (BDNF))

Alternative Name: BDNF

Background: Brain-derived neurotrophic factor, also known as BDNF, is a prosurvival factor induced by cortical neurons that is necessary for survival of striatal neurons in the brain. BDNF is a member of the neurotrophin family of growth factors, which are related to the NGF. It acts on certain neurons of the central nervous system and the peripheral nervous system, helping to support the survival of existing neurons, and encourage the growth and differentiation of new neurons and synapses. In the brain, it is active in the hippocampus, cortex, and basal forebrain—areas vital to learning, memory, and higher thinking. Post mortem analysis has shown lowered levels of BDNF in the brain tissues of people with Alzheimer's disease.

Pathways: RTK Signaling, Synaptic Membrane, Feeding Behaviour, Dicarboxylic Acid Transport, Regulation of long-term Neuronal Synaptic Plasticity