BHLH99 Antibody
Description
BHLH99 Protein Overview
BHLH99 (Basic Helix-Loop-Helix 99) is a plant-specific transcription factor implicated in disease susceptibility to necrotrophic fungal pathogens like Botrytis cinerea and Plectosphaerella cucumerina . Key features include:
-
Structure: Contains a conserved bHLH DNA-binding domain.
-
Function: Regulates expression of PROVIR (provirulence) factors, which enhance host susceptibility to fungal infections .
-
Mechanism: Mutants lacking BHLH99 show hyperactivation of jasmonic acid (JA)-mediated defenses but paradoxically increased disease vulnerability due to upregulated PROVIR genes .
Antibody Development and Validation
While no direct data on BHLH99 antibody production exists, established workflows for antibody characterization (e.g., NeuroMab, CPTAC) provide a framework :
Key Validation Steps
For BHLH99, validation would likely involve Arabidopsis thaliana mutants to confirm absence of signal in bhlh99 knockouts .
Research Applications
BHLH99 antibodies could enable:
-
Pathogen Interaction Studies: Mapping BHLH99’s role in PROVIR gene activation during fungal infections .
-
Subcellular Localization: Determining if BHLH99 localizes to nuclei or interacts with membranes (e.g., via immunofluorescence) .
-
Protein Complex Analysis: Identifying binding partners through co-immunoprecipitation .
Challenges and Considerations
-
Cross-Reactivity: Antibodies targeting bHLH domains must distinguish between closely related family members (e.g., BHLH22 in humans) .
-
Antigen Design: Recombinant BHLH99 protein fragments or synthetic peptides are typical immunogens .
-
Database Gaps: No BHLH99-specific entries exist in major antibody databases (e.g., Antibodypedia, Histone Antibody Specificity Database) .
Future Directions
-
High-Throughput Screening: Adapt NeuroMab’s pipeline (~1,000 clones screened via ELISA/Western blot) for BHLH99.
-
Open-Access Resources: Follow NeuroMab’s model of sharing sequences and protocols to accelerate community validation .
-
Therapeutic Potential: Engineered VHH antibodies (single-domain) could improve tissue penetration in plant studies .
Product Specs
Composition: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
Q&A
Frequently Asked Questions for BHLH99 Antibody Research
(Note: No direct references to "BHLH99" were identified in the provided literature. The below FAQs synthesize general antibody research methodologies from the sources, adapted to hypothetical BHLH99 studies.)
What validation strategies ensure specificity of BHLH99 antibodies in immunohistochemistry (IHC)?
-
Methodological framework:
-
Negative controls: Use tissues/cell lines with confirmed absence of the target antigen (e.g., CRISPR-Cas9 knockouts) .
-
Orthogonal validation: Pair IHC with immunoblotting (reducing vs. non-reducing conditions) and immunoprecipitation-mass spectrometry .
-
Competition assays: Pre-incubate antibodies with recombinant antigen to confirm signal loss .
-
How do experimental designs differ when targeting membrane-bound vs. intracellular antigens with BHLH99?
-
Membrane antigens:
-
Intracellular antigens:
How can conflicting binding data for BHLH99 across studies be resolved?
-
Systematic analysis:
What in silico tools complement empirical validation of BHLH99’s cross-reactivity?
-
Computational pipeline:
How do bispecific antibody platforms influence BHLH99 therapeutic development?
-
Case study framework:
