Customize FRB1 Antibody

Description

Definition and Biological Context of FRB1 Antibody

The FRB1 gene encodes a Golgi-localized, plant-specific protein with a DUF246 domain, critical for maintaining cell adhesion and proper tissue organization in Arabidopsis . The FRB1 Antibody is used to detect and study the localization, expression, and functional interactions of the FRB1 protein. Unlike traditional cell adhesion mutants that exhibit reduced pectin levels, frb1 mutants show altered oligosaccharide composition and pectin methylesterification, suggesting a distinct mechanism of action .

Research Findings and Applications

The FRB1 Antibody has been instrumental in elucidating the molecular role of FRB1 in plant cell adhesion. Key discoveries include:

Role in Cell Wall Architecture

Pectin Methylesterification: FRB1 regulates pectin methylesterification, affecting cell adhesion strength. frb1 mutants exhibit altered pectin dynamics, leading to tissue dissociation and ectopic cell separation .
Oligosaccharide Composition: FRB1 influences galactose- and arabinose-containing oligosaccharides in the Golgi, impacting cell wall extensins and xyloglucan microstructure .

Subcellular Localization

Immunolocalization studies using the FRB1 Antibody confirm its Golgi localization, supporting its role in modifying cell wall components during secretion .

Functional Interactions

While direct protein interactions remain understudied, FRB1 is hypothesized to modulate glycosylation of arabinogalactan proteins (AGPs), which are critical for cell adhesion .

Experimental Applications

The FRB1 Antibody enables precise analysis of FRB1 protein dynamics:

Technique	Purpose
Western Blotting	Quantify FRB1 protein levels in wild-type vs. frb1 mutants
Immunofluorescence	Visualize FRB1 localization in Golgi bodies and cell membranes
Immunoprecipitation	Identify FRB1-interacting proteins (e.g., glycosyltransferases)

Comparative Analysis with Other Cell Adhesion Mutants

Mutant	Pectin Levels	Oligosaccharide Changes	Cell Adhesion Defect
frb1	Normal	Altered galactose/arabinose	Severe dissociation
qua1 (pectin-deficient)	Reduced	N/A	Moderate dissociation

Data source:

Future Directions

The FRB1 Antibody remains essential for:

Protein Interaction Studies: Identifying FRB1’s role in glycosylation pathways.
Agricultural Applications: Engineering crops with enhanced cell adhesion for improved mechanical strength.
Structural Biology: Elucidating FRB1’s 3D conformation using antibody-based crystallization approaches .

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

FRB1 antibody; OFUT33 antibody; At5g01100 antibody; F7J8.80Protein FRIABLE 1 antibody; EC 2.4.1.- antibody; O-fucosyltransferase 33 antibody; O-FucT-33 antibody; O-fucosyltransferase family protein antibody

Target Names

FRB1

Uniprot No.

Q9LFC6

Target Background

Function

FRB1 Antibody targets a glycosyltransferase essential for normal cell adhesion and cell wall integrity.

Database Links

KEGG: ath:AT5G01100

STRING: 3702.AT5G01100.1

UniGene: At.33964

Protein Families

Glycosyltransferase GT65R family

Subcellular Location

Golgi apparatus membrane; Single-pass type II membrane protein.

Tissue Specificity

Ubiquitous. Strong expression in young seedlings, particularly at the junction between hypocotyl and root, in emerging cotyledons, and in parts of the roots. Also detected in the inflorescence (sepals, petals, mature pollen and siliques) and rosette leave

Q&A

FAQs for FRB1 Antibody Research

Advanced Research Questions

How to resolve contradictions in FRB1 localization data across studies?
- Hypothesis-driven approach:
  - Compare extraction protocols (e.g., ammonium oxalate vs. harsh acid treatments) that alter epitope accessibility (Figure 6A) .
  - Quantify FRB1 abundance in Golgi-enriched fractions using subcellular proteomics paired with antibody staining .
Can FRB1 antibodies differentiate between pectin methylesterification states?
- Combine FRB1 immunolabeling with enzymatic treatments (e.g., pectin methylesterase) and quantify shifts in antibody binding intensity using fluorescence microscopy .
- Validate via comparative analysis of uronic acid content and methylesterification levels in frb1 mutants (Table 1) .
How to design co-localization studies for FRB1 and extensin/AGP markers?
- Use multiplex immunofluorescence with FRB1 antibodies and LM1 (extensin) or LM6 (arabinan) antibodies.
- Quantify overlap using Pearson’s correlation coefficient in cortical cell regions (Figure 7D–K) .

Data Analysis & Technical Challenges

Interpreting arabinose/galactose variability in FRB1 mutant cell walls (Table 1):
- Method: Compare neutral sugar profiles across fractions (CDTA, Na₂CO₃, KOH).
- Key finding: frb1 mutants show 50–80% higher arabinose in insoluble fractions, suggesting altered RGI-pectin/AGP crosslinking .
Optimizing FRB1 antibody dilution for quantitative Western blotting:
- Perform checkerboard titrations against serial dilutions of Arabidopsis protein extracts.
- Validate using a recombinant FRB1 standard curve (e.g., 0.1–10 ng/µL) to ensure linear detection range .

Table 1: Neutral Sugar Composition in FRB1 Mutants

Fraction	Genotype	Arabinose (mol%)	Galactose (mol%)	Uronic Acid (µg/mg)
CDTA	FRB1	20.1 ± 0.6	57.3 ± 1.3	47.1 ± 7.6
CDTA	frb1	24.3–25.7 ± 0.9–3.1	51.1–54.4 ± 1.7–3.4	40.8–42.6 ± 3.8–12.6
4M KOH	FRB1	9.3 ± 1.1	22.0 ± 1.0	–
4M KOH	frb1	13.3–15.4 ± 0.6–1.4	25.3–30.2 ± 0.5–1.8	–
Data from 10-day-old seedlings; bold values indicate significant differences (p < 0.05) .

Methodological Recommendations

For epitope mapping: Use recombinant FRB1 domains (e.g., DUF246) to identify antibody-binding regions .
In plantae validation: Generate CRISPR-Cas9 frb1 mutants and correlate antibody signal loss with phenotypic severity (e.g., cell dissociation) .

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FRB1 Antibody