Customize AHL18 Antibody

Description

Definition and Target Protein Overview

AHL18 Antibody is a polyclonal antibody targeting AT-Hook Motif Nuclear Localized Protein 18 (AHL18), a nuclear protein encoded by the AHL18 gene (At3g60870) in Arabidopsis thaliana. This antibody is primarily used in plant biology research to study AHL18’s role in root development, cell proliferation, and organogenesis .

AHL18 Protein Function

AHL18 is part of the AT-hook motif protein family, which regulates chromatin structure and gene expression. Key functional insights include:

Root Apical Meristem Regulation: AHL18 modulates cell proliferation and differentiation in the root apical meristem (RAM), influencing root growth rates .
Lateral Root Development: AHL18 promotes lateral root primordia (LRP) initiation and emergence, with mutants showing 50.7% arrested/delayed LRPs versus 25.2% in wild-type plants .
Cell Cycle Modulation: Overexpression of AHL18 increases S-phase nuclei (by 37.7%), while knockout reduces proliferation (by 20.4%) .

Root System Architecture

Primary Root Growth: ahl18 mutants exhibit shorter primary roots (RAM length reduced by 30%) compared to wild type .
Lateral Root Density: Overexpression lines show a 2.1-fold increase in lateral root initiation events .

Molecular Mechanisms

Gene Expression: AHL18 binds chromatin to regulate genes involved in auxin signaling and cell cycle progression .
Epigenetic Regulation: Phosphoproteomic studies link AHL18 to chromatin remodeling, influencing stress-response pathways .

Experimental Applications

AHL18 Antibody has been utilized in:

Promoter Activity Analysis: Localized AHL18 expression in RAM, differentiating xylem, and pericycle cells using pAHL18::mCherry and pAHL18::GUS lines .
EdU Labeling: Quantified S-phase nuclei in root meristems to assess proliferation rates .
Mutant Phenotyping: Characterized ahl18 knockout lines for reduced organogenesis and delayed lateral root emergence .

Comparative Insights Across Studies

Study Focus	Key Outcome	Citation
Root Meristem Activity	AHL18 overexpression increases RAM length by 45% versus wild type
Lateral Root Initiation	ahl18 mutants show 50% arrested LRPs in branching zones
Cell Cycle Regulation	AHL18 modulates cyclin-dependent kinase activity in pericycle cells

Future Research Directions

Stress Adaptation: Investigate AHL18’s role in drought or nutrient-deficient conditions .
Protein Interactions: Identify AHL18-binding partners using co-immunoprecipitation assays .

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

AHL18 antibody; At3g60870 antibody; T4C21_280 antibody; AT-hook motif nuclear-localized protein 18 antibody

Target Names

AHL18

Uniprot No.

Q9LZX7

Target Background

Function

AHL18 Antibody targets a transcription factor that specifically binds to AT-rich DNA sequences associated with nuclear matrix attachment regions (MARs). It functions redundantly with AHL22, AHL27, and AHL29 in regulating flowering and controlling hypocotyl elongation.

Database Links

KEGG: ath:AT3G60870

STRING: 3702.AT3G60870.1

Subcellular Location

Nucleus.

Q&A

Basic Research Questions

How to validate AHL18 antibody specificity in plant protein studies?
- Methodology:
  - Perform immunoblotting with Arabidopsis thaliana wild-type and AHL18-knockout mutants to confirm absence of signal in mutants .
  - Use peptide blocking assays with the immunogen peptide to verify epitope specificity .
  - Validate across applications (e.g., ICC-IF, IHC) using standardized protocols from repositories like the DSHB .
- Data-driven recommendations:
  - Include a negative control table:
  Sample Type Expected Signal (Wild-Type) Expected Signal (Knockout)
  Western Blot ~55 kDa band No band
  ICC-IF Nuclear/cytoplasmic staining No staining
What experimental applications are supported for AHL18 antibody?
- Methodology:
  - Prioritize immunohistochemistry (IHC) for tissue-specific localization in Arabidopsis roots/shoots .
  - Use immunoprecipitation (IP) paired with mass spectrometry to identify AHL18 interaction partners .
  - Validate in native PAGE to confirm recognition of non-denatured protein conformations .

Sample Type	Expected Signal (Wild-Type)	Expected Signal (Knockout)
Western Blot	~55 kDa band	No band
ICC-IF	Nuclear/cytoplasmic staining	No staining

Advanced Research Questions

How to resolve discrepancies in AHL18 subcellular localization across studies?
- Analysis framework:
  - Compare fixation methods: paraformaldehyde (preserves membrane integrity) vs. methanol (denatures epitopes) .
  - Use live-cell imaging with fluorescently tagged AHL18 as a reference .
- Data contradiction resolution:
  - Example troubleshooting table:
  Fixation Method Localization Reported Likelihood of Artifact
  Methanol Cytoplasmic High (epitope masking)
  Paraformaldehyde Nuclear Low
How to optimize AHL18 antibody for non-model plant species?
- Methodology:
  - Perform epitope mapping to identify conserved regions across species using tools like Clustal Omega .
  - Test cross-reactivity with protein extracts from closely related species (e.g., Brassica napus) via Western Blot .
- Recommendations:
  - If cross-reactivity is low, consider recombinant antibody engineering to enhance affinity for divergent epitopes .
How to address cross-reactivity with other AHL family proteins (e.g., AHL2, AHL26)?
- Experimental design:
  - Use ELISA-based competition assays with purified AHL2/AHL26 proteins to quantify cross-binding .
  - Generate a structural alignment of AHL family epitopes to identify unique residues in AHL18 .
- Data interpretation:
  - Cross-reactivity profile example:
  Protein Tested % Signal Reduction (vs. AHL18)
  AHL2 15%
  AHL26 8%
  Non-target 95%

Fixation Method	Localization Reported	Likelihood of Artifact
Methanol	Cytoplasmic	High (epitope masking)
Paraformaldehyde	Nuclear	Low

Protein Tested	% Signal Reduction (vs. AHL18)
AHL2	15%
AHL26	8%
Non-target	95%

Methodological Best Practices

For reproducibility:
- Use recombinant AHL18 antibodies when available to minimize batch variability .
- Deposit validation data in public repositories (e.g., Antibody Registry) to aid community verification .
For troubleshooting:
- If signal is weak in IHC, employ antigen retrieval with citrate buffer (pH 6.0) to expose epitopes .
- For quantitative assays, normalize signals using housekeeping proteins validated in Arabidopsis (e.g., ACTIN2) .

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AHL18 Antibody