Customize SMO1-3 Antibody

Description

Biological Context of SMO1-3

SMO1-3 is one of three Arabidopsis thaliana SMO1 isoforms (SMO1-1, SMO1-2, and SMO1-3) that catalyze the oxidation of 4α-methylsterols during phytosterol biosynthesis . These enzymes are essential for:

Embryo development: Double mutants (smo1-1 smo1-2) exhibit severe embryonic lethality due to disrupted auxin and cytokinin signaling .
Sterol homeostasis: SMO1 enzymes regulate cholesterol-like compounds critical for membrane integrity and hormone signaling .
Auxin-cytokinin crosstalk: Impaired SMO1 activity leads to elevated auxin responses and reduced cytokinin activity, perturbing cell differentiation .

Table 1: Functional Roles of SMO1 Isoforms

Isoform	Mutant Phenotype	Key Function
SMO1-1	Lethal in homozygous smo1-1 smo1-2 mutants	Catalyzes sterol oxidation; maintains auxin-cytokinin balance
SMO1-2	Embryonic lethality in double mutants	Required for sterol-dependent signaling pathways
SMO1-3	Partially compensates for SMO1-1/SMO1-2 loss	Redundant role in sterol metabolism

**Table 2: Impact of smo1 Mutants on Hormonal Pathways**

Parameter	Wild-Type	smo1-1/+ smo1-2 Mutants
Auxin response	Baseline	↑ 2–3× (e.g., DR5rev:GFP expression)
Cytokinin biosynthesis	Normal	↓ 50–70% (e.g., IPT3 expression)
Callus formation	Moderate	Accelerated at low cytokinin concentrations

Mechanistic Insights

Sterol binding to Smoothened (SMO): While unrelated to the Hedgehog pathway’s SMO protein in animals , plant SMO1-3 indirectly influences lipid-mediated signaling by modulating sterol availability .
Gene redundancy: Single smo1-3 mutants show no visible defects, but combinatorial knockouts (e.g., smo1-1 smo1-2) are lethal, highlighting functional overlap .
Therapeutic potential: Antibodies targeting sterol-metabolizing enzymes (e.g., SMO1-3) could inform strategies for engineering stress-resistant crops .

Antibody Validation and Applications

Though the provided sources lack direct data on SMO1-3 antibody validation, analogous studies on plant antibodies emphasize:

Western blot: Expected band size ~86 kDa (based on SMO1-3’s calculated molecular weight) .
Immunolocalization: Used to track SMO1-3 expression in embryonic tissues and meristems .
Functional assays: Critical for quantifying sterol levels in smo1 mutants via ELISA or LC-MS .

Product Specs

Buffer

Preservative: 0.03% ProClin 300; Constituents: 50% Glycerol, 0.01M PBS, pH 7.4

Form

Liquid

Lead Time

14-16 weeks (made-to-order)

Synonyms

SMO1-3; At4g22755; T12H17.140; Methylsterol monooxygenase 1-3; Sterol 4-alpha-methyl-oxidase 1-3; AtSMO1-3

Target Names

SMO1-3

Uniprot No.

F4JLZ6

Target Background

Function

SMO1-3 Antibody targets a non-heme iron oxygenase involved in sterol biosynthesis. The preferred substrates are 4,4-dimethyl-9β,19-cyclopropylsterols, such as 24-methylenecycloartanol.

Database Links

KEGG: ath:AT4G22755

Protein Families

Sterol desaturase family

Subcellular Location

Endoplasmic reticulum membrane; Multi-pass membrane protein.

Q&A

Here’s a structured FAQ collection for researchers working with SMO1-3 antibodies, optimized for academic rigor and methodological depth:

Advanced Research Questions

How to design a study investigating SMO1-3’s role in non-canonical RAS pathways?

Framework:
- Genetic Models: Use CRISPR-edited cells with SMO1-3 KO + inducible rescue mutants.
- Functional Assays: Measure RAS activation via FRET biosensors under ligand-stimulated vs. basal conditions .
- Proteomic Integration: Combine IP-MS with phosphoproteomics to map novel interactors .
- Key Control: Include cells expressing conformation-locked ErbB3 mutants to isolate SMO1-3-specific effects .

How to resolve contradictory data on SMO1-3’s binding affinity in structural studies?

Analysis Workflow:

Technique Comparison:

Method	Affinity Range	Strengths	Limitations
Surface Plasmon Resonance (SPR)	pM–nM	Quantifies monovalent binding kinetics	Requires purified protein
Flow Cytometry	nM–μM	Measures cell-surface binding in native state	Limited by epitope accessibility

Variable Control: Account for glycosylation states (e.g., deglycosylate samples using PNGase F) .

What advanced techniques enable single-cell resolution of SMO1-3 dynamics?

Emerging Methods:
- CITE-seq: Pair antibody-derived tags with transcriptomics to correlate protein localization with gene expression .
- Live-Cell Imaging: Use pH-sensitive GFP fusions to track real-time antibody internalization .
- Nanobody Engineering: Develop camelid-derived nanobodies for improved tissue penetration (e.g., PRL-3-targeting alpaca antibodies) .

Data Contradiction Resolution

How to address discrepancies in post-translational modification detection?

Stepwise Approach:
- Middle-Level LC-MS: Use FabRICATOR-digested fragments (23–25 kDa) for glycosylation profiling .
- Redox-State Testing: Compare reducing vs. non-reducing SDS-PAGE to identify disulfide bond artifacts .
- Temporal Analysis: Sample at multiple timepoints (0–24h post-stimulation) to capture dynamic modifications .

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SMO1-3 Antibody