Customize H0219H12.8 Antibody

Product Specs

Buffer

Preservative: 0.03% Proclin 300
Constituents: 50% Glycerol, 0.01M Phosphate Buffered Saline (PBS), pH 7.4

Form

Liquid

Lead Time

Made-to-order (14-16 weeks)

Synonyms

H0219H12.8; OsI_16169; OSIGBa0097P08.13; Casparian strip membrane protein 6; OsCASP6

Target Names

H0219H12.8

Uniprot No.

A2XU91

Target Background

Function

This antibody targets a protein involved in the regulation of membrane-cell wall junctions and localized cell wall deposition. It is essential for the formation of the Casparian strip membrane domain (CSD) and subsequent development of Casparian strips. These specialized cell wall modifications in the root endodermis act as a barrier, preventing lateral diffusion between the apoplasm within the organism and the apoplasm outside the organism.

Protein Families

Casparian strip membrane proteins (CASP) family

Subcellular Location

Cell membrane; Multi-pass membrane protein.

Q&A

What is the primary target and biological relevance of the H0219H12.8 Antibody?

The H0219H12.8 Antibody targets the Casparian strip membrane protein OsI_16169 (H0219H12.8) in Oryza sativa subsp. indica (rice). This protein is implicated in root tissue structure and function, particularly in forming apoplastic barriers that regulate ion transport and nutrient uptake . Its study is critical for understanding plant responses to abiotic stressors like drought .

Methodological Note:
For initial validation, use Western blotting with recombinant OsI_16169 protein (e.g., the ELISA recombinant protein product ) to confirm antibody specificity. Include negative controls (e.g., non-specific IgG) to rule out cross-reactivity .

How do I validate antibody specificity for H0219H12.8 in plant tissues?

Validation requires a multi-step approach:

Method	Protocol	Purpose
Western Blot	1. Extract total protein from rice root tissues. 2. Resolve via SDS-PAGE. 3. Transfer to PVDF membrane. 4. Incubate with H0219H12.8 Antibody (1:1000 dilution). 5. Use HRP-conjugated secondary antibody for detection.	Confirm antibody binds to a single band (~21 kDa, predicted size for OsI_16169 ).
Immunoprecipitation	1. Lyse plant cells with RIPA buffer. 2. Incubate lysate with antibody-bound beads. 3. Elute bound proteins. 4. Analyze via Western blot.	Verify antibody binds native protein and excludes non-specific binding.
ELISA	1. Coat plates with recombinant OsI_16169 protein . 2. Perform sandwich ELISA with H0219H12.8 Antibody. 3. Use non-specific IgG as negative control.	Quantify antibody affinity (e.g., EC₅₀ values).

Key Consideration: Cross-reactivity with homologous proteins (e.g., other Casparian strip proteins) should be assessed using peptide competition assays .

What experimental controls should I include when using this antibody?

Critical controls include:

Negative Control: Non-specific IgG (e.g., rabbit IgG) to detect background signal.
Positive Control: Recombinant OsI_16169 protein to confirm antibody functionality.
No Primary Antibody Control: Exclude secondary antibody interactions.
Tissue-Specific Controls: Compare root vs. shoot tissues to confirm spatial expression patterns .

How do I address potential cross-reactivity in complex plant lysates?

Cross-reactivity risks arise due to conserved epitopes in plant membrane proteins. Mitigation strategies:

Approach	Method	Outcome
Epitope Mapping	1. Use synthetic peptides spanning OsI_16169 sequence. 2. Perform dot blot or ELISA.	Identify antibody-binding regions to design blocking peptides.
Blocking Agents	1. Pre-incubate antibody with excess non-specific proteins (e.g., BSA). 2. Add blocking peptides.	Reduce non-specific binding to abundant plant proteins like rubisco.
Computational Prediction	1. Use tools like BlastP to identify homologous proteins. 2. Design siRNA/RNAi to knock down targets.	Validate antibody specificity via gene-silencing experiments .

How does computational modeling inform H0219H12.8 Antibody optimization?

Computational tools enhance antibody design and validation:

Tool	Application	Reference
Homology Modeling	Predict 3D structure of OsI_16169 to identify surface-exposed epitopes.	PMC7947987
Docking	Simulate antibody-antigen interactions to optimize binding affinity.	PMC7947987
B Cell Receptor Repertoire Analysis	Map antibody sequences to identify clonal diversity.	Enables selection of high-affinity clones.

Case Study: In drought-responsive gene studies, computational tools help prioritize antibodies targeting stress-responsive proteins like OsI_16169 .

What advanced techniques can I use to study OsI_16169 function in vivo?

For functional studies, integrate:

CRISPR Knockout/Knock-In:
- Generate osI_16169 mutants to study phenotypic effects (e.g., altered root permeability).
- Use H0219H12.8 Antibody to confirm protein absence/presence.
Proximity Ligation Assay (PLA):
- Detect protein-protein interactions (e.g., OsI_16169 with transporters) in fixed plant tissues.
Single-Molecule Localization Microscopy (SMLM):
- Map OsI_16169 localization at super-resolution to study Casparian strip dynamics.

Why might I observe weak/no signal in Western blotting?

Common issues and solutions:

Problem	Cause	Solution
Low Signal	Insufficient antibody concentration.	Optimize dilution (e.g., 1:500 to 1:2000).
High Background	Non-specific binding.	Use blocking agents (e.g., 5% BSA) or pre-clear lysate.
Protein Degradation	Inactive proteases in lysate.	Add protease inhibitors (e.g., PMSF).

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H0219H12.8 Antibody