Customize At3g14380 Antibody

Description

Immunogen and Reactivity

The immunogen is a recombinant protein corresponding to the At3g14380 gene product. The antibody exhibits high specificity for Arabidopsis thaliana samples, validated through ELISA and WB . Its polyclonal nature ensures recognition of multiple epitopes, enhancing detection sensitivity.

Applications in Chromatin Research

The At3g14380 gene encodes an integral membrane protein implicated in chromatin organization. Research on related Arabidopsis MORC (Microrchidia) proteins, such as AtMORC1 and AtMORC6, reveals their role in chromocenter condensation and heterochromatin compaction. Mutants lacking these proteins show decondensed chromocenters and altered H3K9me2 distribution, though At3g14380 itself is not directly linked to histone modification . The At3g14380 antibody enables targeted studies of this protein’s localization and interaction networks in plant nuclei.

Key Research Findings

Chromocenter Dynamics: AtMORC proteins, including those encoded by genes like At3g14380, are critical for maintaining heterochromatin integrity. Mutants exhibit disrupted chromocenter condensation, as visualized via H3K9me2 immunostaining .
Epigenetic Regulation: While At3g14380 mutants do not alter H3K9me2 levels globally, they may influence transcriptional silencing at specific loci .

Biological Significance

The At3g14380 protein is part of a conserved family of chromatin-associated ATPases. Its study contributes to understanding plant epigenetic mechanisms, particularly in stress responses and development. The antibody’s utility in detecting this protein supports investigations into nuclear architecture and gene silencing pathways in Arabidopsis .

For extended use, pair with secondary antibodies such as alkaline phosphatase–conjugated goat anti-rabbit IgG (H+L) (e.g., Product SA00002-2) .

Product Specs

Buffer

**Preservative:** 0.03% Proclin 300
**Constituents:** 50% Glycerol, 0.01M PBS, pH 7.4

Form

Liquid

Lead Time

Made-to-order (14-16 weeks)

Synonyms

At3g14380; MLN21.16; CASP-like protein 2A2; AtCASPL2A2

Target Names

At3g14380

Uniprot No.

Q9LUL1

Target Background

Function

This antibody targets a protein that plays a role in the shedding of floral organs.

Database Links

KEGG: ath:AT3G14380

STRING: 3702.AT3G14380.1

UniGene: At.39246

Protein Families

Casparian strip membrane proteins (CASP) family

Subcellular Location

Cell membrane; Multi-pass membrane protein.

Tissue Specificity

Mostly expressed in flowers and buds and, to a lower extent, in roots and yellow siliques. Localized in the floral organ abscission zone.

Q&A

Here’s a structured FAQ collection for researchers working with the At3g14380 antibody, optimized for academic rigor and methodological depth:

Advanced Research Questions

How do I resolve conflicting WB results between At3g14380 antibody and RNA-seq data?
- Step 1: Confirm antibody specificity via siRNA knockdown or CRISPR-Cas9 knockout lines.
- Step 2: Check for post-translational modifications (e.g., phosphorylation) that may alter epitope accessibility .
- Step 3: Quantify protein vs. transcript levels using parallel MRM-MS (mass spectrometry) and qRT-PCR .
What experimental designs minimize aggregation during immunoprecipitation (IP)?
- Buffer optimization: Use 25 mM HEPES (pH 7.4), 150 mM NaCl, and 0.1% Tween-20 to reduce non-specific interactions .
- Antibody:bead ratio: Pre-test ratios (e.g., 1:2 to 1:10) to balance signal and background .
- Controls: Include IgG isotype controls and “no-antibody” beads .

How does At3g14380 antibody performance compare to epitope-tagged alternatives (e.g., GFP-fusion)?

Parameter	At3g14380 Antibody	GFP-Tagged Systems
Native conformation	Yes	May alter protein folding
Quantification	Semi-quantitative (WB)	Quantitative (fluorescence)
Cross-reactivity risk	Low (species-specific)	High (GFP antibodies)

Data Interpretation & Troubleshooting

Why does ELISA show higher sensitivity than WB for At3g14380 detection?
- ELISA captures linear epitopes (denatured), while WB may miss conformational epitopes . Optimize denaturation conditions (e.g., 95°C for 5 min with β-mercaptoethanol).
How to address background noise in immunofluorescence (IF)?
- Pre-absorption: Incubate antibody with Arabidopsis leaf lysate (non-transgenic) for 1 hr .
- Secondary antibody validation: Use anti-rabbit IgG pre-adsorbed against plant proteins .

Methodological Innovations

Can this antibody be engineered for improved pH stability in live-cell imaging?
- Apply CH3 domain mutations (e.g., N392K/M397V) to enhance thermal stability. Validate via differential scanning calorimetry (DSC) .
What computational tools predict At3g14380-antibody binding interfaces?
- Use ClusPro 2.0 for docking simulations or AlphaFold-Multimer for epitope mapping . Pair with mutagenesis (e.g., alanine scanning) to confirm critical residues.

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At3g14380 Antibody