Customize AHL16 Antibody

Description

Overview of ARL16 Antibodies

ARL16 antibodies are immunodetection tools targeting the ADP-ribosylation factor-like 16 protein, a small GTPase implicated in intracellular trafficking and ciliary function . These antibodies are widely used in techniques such as Western blotting (WB), immunohistochemistry (IHC), and enzyme-linked immunosorbent assays (ELISA) .

Role in Ciliogenesis

ARL16 regulates ciliary protein trafficking, particularly in the export of IFT140 (intraflagellar transport protein) and INPP5E (inositol polyphosphate phosphatase) from the Golgi to cilia. Knockout (KO) of Arl16 in mouse embryonic fibroblasts (MEFs) results in:

Reduced ciliogenesis (fewer ciliated cells) .
Elongated cilia in remaining cells .
Accumulation of IFT140 and INPP5E at the Golgi, indicating disrupted trafficking .

Subcellular Localization

In human retinal pigmented epithelial (RPE1) cells, ARL16 localizes to:

Ciliary axonemes (punctate staining) .
Mitochondria (colocalization with HSP60) .
Cytosol (diffuse distribution) .

Validation Data Examples

Supplier	Validation Method	Result
Antibodies.com	ELISA, WB, IHC	Confirmed reactivity in human samples
Sigma-Aldrich	Prestige Antibodies®	Atlas-based staining in 44 human tissues
Thermo Fisher	Ortholog cross-reactivity	80% sequence homology in mouse

Limitations and Considerations

Species Specificity: Most ARL16 antibodies are optimized for human samples, with limited reactivity in rodents .
Localization Challenges: Commercially available antibodies fail to detect endogenous ARL16 in murine models due to low epitope conservation .
Functional Studies: No small-molecule inhibitors or activators of ARL16 are reported; genetic KO remains the primary functional analysis tool .

Future Directions

Research priorities include:

Developing ARL16-specific modulators to dissect its GTPase activity.
Investigating ARL16’s role in mitochondrial dynamics and cilia-related pathologies (e.g., retinal degeneration).
Expanding antibody validation to 3D organoid models for disease relevance .

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

AHL16 antibody; TEK antibody; At2g42940 antibody; F23E6.8 antibody; F7D19.6AT-hook motif nuclear-localized protein 16 antibody; Protein TRANSPOSABLE ELEMENT SILENCING VIA AT-HOOK antibody; Protein TEK antibody

Target Names

AHL16

Uniprot No.

Q9SJG4

Target Background

Function

AHL16 is a transcription factor exhibiting specific binding affinity for AT-rich DNA sequences associated with nuclear matrix attachment regions (MARs). It encodes a nuclear matrix protein crucial for maintaining genomic integrity by epigenetically silencing transposable elements (TEs) and repeat-containing genes. Furthermore, AHL16 functions as a chromatin remodeling factor, influencing the architecture of FLC and FWA chromatin through modulation of H3 acetylation and methylation, thereby regulating FLC and FWA expression. It negatively regulates floral repressors, including MAF4 and MAF5, and plays a vital role in activating transcription during anther development. Additionally, AHL16 regulates the expression of arabinogalactan proteins (AGPs) involved in nexine layer formation within the pollen wall. Specifically, it binds to the promoters of AGP6, AGP11, AGP23, and AGP40.

Gene References Into Functions

Studies indicate that MADS AFFECTING FLOWERING 4 (MAF4) and MAF5 are upregulated in plants with reduced AHL16 expression. This demonstrates AHL16's negative regulatory role over these floral repressors. (PMID: 23733063)

Database Links

KEGG: ath:AT2G42940

STRING: 3702.AT2G42940.1

UniGene: At.45674

Subcellular Location

Nucleus.

Tissue Specificity

Preferentially expressed in the inflorescence meristem and young floral buds, as well as in seedling-stage vegetative meristems. Widely expressed in flowers, roots and stems, with relatively low expression in leaves.

Q&A

Here’s a structured collection of FAQs for researchers focused on CD16A antibodies (interpreted as the likely subject, given the absence of "AHL16" in provided sources), addressing academic research scenarios with methodological rigor and data-supported insights:

Advanced Experimental Design & Challenges

How can researchers address immunogenicity risks in CD16A antibody clinical trials?

Preclinical screening: Use HLA-transgenic mouse models to predict T-cell epitopes.
Clinical monitoring: Implement HAHA assays with thresholds >50 ng/mL for intervention .
Engineering strategies: Employ deimmunization algorithms to remove putative immunogenic motifs from variable regions .

What methods resolve contradictions in antibody specificity data across studies?

Adopt the "five pillars" of antibody validation:

Genetic controls: Use CRISPR-Cas9 KO cell lines to confirm on-target effects .
Orthogonal validation: Pair flow cytometry with immunohistochemistry in CD16A+ tissues.
Independent antibodies: Compare results from ≥2 clones (e.g., clone 3G8 vs. LNK16) .
Immunocapture-MS: Identify off-target proteins co-precipitated by the antibody .

How do allotype-specific differences (e.g., FcγRIIIa-158V/F) impact antibody development?

Binding variance: The 158V variant binds IgG1 with ~10-fold higher affinity than 158F, affecting ADCC efficacy .
Design solutions: Develop allotype-independent antibodies by targeting conserved epitopes (e.g., membrane-proximal domains) .

Data Conflict Resolution Table

Conflict Type	Resolution Strategy	Example
Immunogenicity discrepancies (humanized vs. chimeric)	Retrospective HAHA analysis across trials	CDR-grafted A33 antibody showed higher immunogenicity (45% HAHA+) vs. chimeric B72.3 (12% HAHA+) .
Off-target binding in flow cytometry	Use KO controls and titrate antibodies	False positivity in CD16A staining resolved by KO validation and optimizing concentrations to ≤1 μg/mL .

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AHL16 Antibody