BDNF Antibody
Description
Definition and Types of BDNF Antibodies
BDNF antibodies are immunoglobulin-based reagents that bind specifically to BDNF or its precursor (proBDNF). They are classified into two broad categories:
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Polyclonal antibodies (e.g., Rabbit anti-Human BDNF from Bio-Rad ): Recognize multiple epitopes, offering high sensitivity but potential cross-reactivity.
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Monoclonal antibodies (e.g., Clone EPR1292 from Abcam ): Target a single epitope, ensuring specificity and batch-to-batch consistency.
Biomarker Analysis
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Major Depressive Disorder (MDD): Serum mature BDNF levels are significantly reduced in MDD patients (21.09 ± 5.60 ng/mL vs. 23.11 ± 5.90 ng/mL in controls) .
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Bipolar Disorder (BP): BDNF levels correlate with disease progression, showing lower levels in late-stage BP .
Mechanistic Studies
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Chronic Pain: BDNF-neutralizing antibody B30 reverses hyperexcitability in sensory neurons, demonstrating 30-fold higher potency than its parent antibody in nerve injury models .
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Synaptic Plasticity: BDNF antibodies validate BDNF’s role in long-term potentiation (LTP) and depression (LTD) via TrkB receptor modulation .
Clinical and Therapeutic Implications
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Diagnostic Potential: BDNF antibodies are used in ELISA kits to quantify serum BDNF, aiding in stratification of psychiatric disorders .
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Therapeutic Development: High-affinity monoclonal antibodies (e.g., B30) show promise in treating neuropathic pain by sequestering peripheral BDNF without CNS penetration .
Validation Challenges
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Cross-Reactivity: Commercial kits vary widely; Promega-Emax® and Biosensis antibodies detect proBDNF, whereas R&D Systems’ Quantikine® is specific for mature BDNF .
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Assay Reproducibility: Inter-kit variability necessitates standardization, as highlighted by a study comparing six commercial BDNF detection kits .
Recommended Protocols
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Western Blot: Use 1:500–1:1000 dilution for Proteintech’s 25699-1-AP .
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Immunohistochemistry: Optimal concentrations range from 2–5 µg/mL for monoclonal antibodies .
Emerging Innovations
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Affinity-Optimized Antibodies: B30, an engineered variant of R3bH01, shows >300-fold improved affinity and efficacy in reversing nerve injury phenotypes .
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Peripheral Targeting: Antibodies like B30 are designed to avoid CNS penetration, minimizing cognitive side effects during chronic pain treatment .
Product Specs
Q&A
What criteria should guide BDNF antibody selection for Western blotting?
Three key parameters require verification:
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Expected molecular weight recognition: Pro-BDNF migrates at 32-35 kDa while mature BDNF appears at 14-18 kDa . Valid antibodies must demonstrate band patterns matching these predictions in overexpression systems.
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Knockout validation: Test antibodies in BDNF⁻/⁻ tissue lysates to confirm absence of non-specific bands .
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Isoform discrimination: Use recombinant pro-BDNF and mature BDNF in parallel blots (Table 1).
Table 1: Western Blot Performance Characteristics of Common BDNF Antibodies
| Antibody Clone | pro-BDNF Detection | Mature BDNF Detection | KO Validation |
|---|---|---|---|
| R3bH01 | No | Yes (14 kDa) | Confirmed |
| 35928 | Yes (32 kDa) | Weak | Partial |
| Biosensis BEK | No | Yes (18 kDa) | Confirmed |
How should researchers design controls for BDNF immunohistochemistry?
Implement a three-tier control system:
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Biological controls: Compare staining intensity between wild-type and BDNF-deficient tissues .
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Technical controls: Include secondary antibody-only slides and pre-immune serum controls .
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Competition controls: Pre-incubate antibodies with 10x molar excess recombinant BDNF for 1 hr at 37°C . In spinal cord sections, this should reduce signal intensity by >80% in validated antibodies .
How to resolve contradictory BDNF localization data between ELISA and IHC?
Contradictions often arise from:
Epitope accessibility differences:
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ELISA detects both intracellular and secreted BDNF (linear epitopes)
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IHC requires intact tertiary structure (conformational epitopes)
Methodological reconciliation protocol:
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Perform antigen retrieval optimization (pH 9.0 vs pH 6.0 buffers)
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Compare multiple fixation methods (PFA vs methanol)
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Validate with RNAscope® BDNF probes as orthogonal verification
How to validate BDNF-neutralizing antibodies for functional studies?
A three-assay validation cascade is essential:
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Surface plasmon resonance: Confirm Kd < 1 nM for therapeutic applications
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pERK inhibition assay: Measure IC50 in TrkB-expressing HEK293 cells (ideal range: 0.1-5 nM)
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Ex vivo electrophysiology: Test reversal of LTP in hippocampal slices at 1-10 μg/ml
Critical data interpretation considerations:
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Neutralizing capacity depends on antibody:BDNF stoichiometry (≥2:1 required)
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Biological activity persists for 48-72 hrs post-application in slice cultures
What explains variable BDNF antibody performance across laboratories?
Four key variables require standardization:
How to address discrepancies in BDNF quantification between techniques?
Develop a consensus normalization protocol:
Product Science Overview
Brain-Derived Neurotrophic Factor (BDNF) is a member of the neurotrophin family of growth factors, which are essential for the development, maintenance, and function of the nervous system. BDNF plays a crucial role in neuronal survival, differentiation, and synaptic plasticity, making it a key player in learning, memory, and higher cognitive functions .
BDNF is a protein that is encoded by the BDNF gene. It is initially synthesized as a precursor protein, proBDNF, which is then cleaved to produce the mature BDNF protein. The mature BDNF protein consists of 247 amino acids and forms a homodimer through disulfide bonds. BDNF binds to two types of receptors on the surface of neurons: the high-affinity receptor TrkB (tropomyosin receptor kinase B) and the low-affinity receptor p75NTR .
BDNF is a critical mediator of neuroplasticity, the brain’s ability to reorganize itself by forming new neural connections. This process is essential for learning, memory, and recovery from brain injuries. BDNF promotes the growth and differentiation of new neurons and synapses, and it enhances the strength of existing synapses .
Mouse anti-human BDNF antibodies are monoclonal antibodies produced by immunizing mice with human BDNF protein. These antibodies are highly specific and are used in various research applications, including Western blotting, immunohistochemistry, immunocytochemistry, enzyme-linked immunosorbent assays (ELISA), and flow cytometry .
- Neurodegenerative Diseases: BDNF levels are often altered in neurodegenerative diseases such as Alzheimer’s disease, Parkinson’s disease, and Huntington’s disease. Mouse anti-human BDNF antibodies are used to study the expression and distribution of BDNF in these conditions .
- Psychiatric Disorders: BDNF is also implicated in psychiatric disorders such as depression, schizophrenia, and bipolar disorder. Research using mouse anti-human BDNF antibodies helps to elucidate the role of BDNF in these disorders and to develop potential therapeutic strategies .
- Neurodevelopmental Disorders: BDNF is essential for normal brain development, and its dysregulation is associated with neurodevelopmental disorders such as autism spectrum disorder and Rett syndrome. Mouse anti-human BDNF antibodies are used to investigate the mechanisms underlying these disorders .
