Customize APE1L Antibody

Description

Introduction to APE1L Antibody

APE1L (apurinic/apyrimidinic endonuclease 1-like) antibodies are specialized reagents designed to detect and study the APE1L protein, a multifunctional enzyme critical for DNA repair, redox regulation, and active DNA demethylation. These antibodies enable researchers to investigate APE1L's roles in genomic stability, immune responses, and epigenetic regulation through techniques such as Western blotting (WB), immunohistochemistry (IHC), and flow cytometry .

Key Features of APE1L Antibodies

Epitope and Reactivity

Recognizes full-length APE1/Ref-1 protein .
Confirmed specificity via knockout controls (e.g., loss of signal in APEX1 KO HAP1 cells) .

Research Applications and Findings

3.1 DNA Repair and Demethylation
APE1L antibodies have been pivotal in elucidating the enzyme’s role in:

Active DNA Demethylation: APE1L processes 3′-blocking DNA ends generated by ROS1 and DME glycosylases, facilitating base excision repair (BER) .
Chromatin Dynamics: Co-localizes with ROS1 and ZDP in subnuclear foci, critical for FWA and MEA gene imprinting in Arabidopsis .

Table 1: Key Functional Insights Using APE1L Antibodies

Function	Experimental Evidence	Source
DNA repair	APE1L processes 3′-PUA termini (weak 3′-phosphatase activity) in vitro
Gene imprinting	ape1l/zdp double mutants show embryonic lethality and altered methylation patterns
Immune modulation	APE1L inhibition enhances Th1 responses via IL-12 retention in dendritic cells

Immune System Regulation

APE1L regulates redox-sensitive transcription factors (e.g., NF-κB, STAT3) to modulate cytokine production (e.g., TNFα, IL-6) .
In antigen-presenting cells (APCs), APE1L knockdown increases IL-12 surface retention and p38 MAPK activation, amplifying Th1 immune responses .

Clinical and Diagnostic Relevance

Cancer Research

Overexpressed APE1L correlates with chemotherapy resistance in tumors .
Nuclear staining patterns in cancers (e.g., osteosarcoma, germ cell tumors) highlight its diagnostic potential .

Neuroinflammation and Neurodegeneration

APE1L’s redox activity in the CNS influences neuroinflammatory pathways, making it a therapeutic target for diseases like Alzheimer’s .

Future Directions

Therapeutic Targeting: Small-molecule inhibitors (e.g., E3330) blocking APE1L’s redox function show promise in modulating immune responses .
Epigenetic Studies: APE1L’s role in active DNA demethylation warrants further exploration in developmental biology and cancer epigenetics .

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

APE1L antibody; At3g48425 antibody; T29H11.60DNA-(apurinic or apyrimidinic site) lyase antibody; EC 3.1.-.- antibody; EC 4.2.99.18 antibody; APEX1-like protein antibody; Apurinic-apyrimidinic endonuclease antibody

Target Names

APE1L

Uniprot No.

Q5XF07

Target Background

Function

APE1L (Apurinic/apyrimidinic endonuclease 1-like) is an enzyme involved in active DNA demethylation and gene imprinting. Studies have reported conflicting results regarding its 3'-phosphatase activity; one study demonstrated in vitro activity, while another did not. APE1L catalyzes the conversion of the 3'-blocking group 3'-phosphor-α,β-unsaturated aldehyde (3'-PUA), a product of ROS1 activity, to 3'-OH. It exhibits strong, non-specific DNA affinity. Functional redundancy exists with APE2; at least one functional allele of either APE1L or APE2 is necessary for seed viability.

Gene References Into Functions

APE1L is a crucial component of the active DNA demethylation pathway in Arabidopsis. In conjunction with ZDP, it regulates gene imprinting. (PMID: 25569774)
Studies of Ape1L, Ape2, and Arp heterozygous mutants demonstrate that at least one functional copy of either Ape1L or Ape2 is essential for complete embryo development. (PMID: 19172180)

Database Links

KEGG: ath:AT3G48425

STRING: 3702.AT3G48425.1

UniGene: At.50257

Protein Families

DNA repair enzymes AP/exoA family

Subcellular Location

Nucleus. Nucleus, nucleolus.

Tissue Specificity

Expressed in leaves, flower buds and developing siliques. Not detected in roots.

Q&A

Frequently Asked Questions: APE1L Antibody in Academic Research

Advanced Research Questions

How to resolve contradictory data on APE1L’s interaction with ROS1 in different cell types?

Troubleshooting framework:
- Cell cycle synchronization: APE1L-ROS1 co-localization varies by cell cycle phase (e.g., nucleolar co-localization in 54% of cells during G1) ( ).
- Post-translational modifications: Screen for phosphorylation/acetylation states affecting interaction dynamics (e.g., redox-sensitive cysteine residues) ( ).
- Single-molecule imaging: Use live-cell tracking to monitor real-time interaction kinetics ( ).

What strategies optimize APE1L antibody performance in low-abundance chromatin contexts?

Protocol enhancements:
- Chromatin shearing: Adjust sonication conditions to preserve APE1L-bound DNA-protein complexes for ChIP-seq ( ).
- Signal amplification: Combine tyramide-based amplification with anti-APE1L [EPR18378-45] (ab189474) for low-copy targets ( ).
- Crosslinking: Test formaldehyde vs. DSG for stabilizing transient interactions ( ).

How to address APE1L antibody cross-reactivity with human APE1/Ref-1 in dual-species systems?

Experimental solutions:
- Phylogenetic divergence: Target non-conserved regions (e.g., plant-specific N-terminal residues) ( ).
- Competitive ELISA: Pre-incubate antibodies with human APE1 protein to block cross-reactive epitopes ( ).

Data Interpretation & Technical Challenges

How to reconcile APE1L’s dual roles in DNA repair and transcriptional regulation?

Integrated analysis:
- Functional genomics: Correlate APE1L ChIP-seq peaks with RNA Pol II occupancy and DNA methylation marks ( ).
- Pathway enrichment: Use gene ontology (GO) terms linked to ape1l mutant phenotypes (e.g., seed development, imprinting defects) ( ).

What controls are critical for APE1L activity assays in vitro?

Essential controls:

Control Type	Purpose	Example
Substrate specificity	Rule out non-target cleavage	Compare activity on 3′-PUA vs. 3′-phosphate DNA ( )
Catalytic mutants	Confirm enzyme dependency	Use N224D-APE1L (loss-of-function) ( )
Metal ion dependence	Validate Mg²⁺/Mn²⁺ cofactor requirements	Test activity in EDTA-treated reactions ( )

Emerging Research Directions

How to design multiplex assays for APE1L interaction networks?

Multiplexing strategies:
- Proximity ligation (PLA): Pair anti-APE1L with anti-ZDP/ROS1 to visualize interaction foci ( ).
- CUT&Tag: Profile APE1L-associated chromatin regions alongside ROS1 binding sites ( ).

What bioinformatic tools decode APE1L’s structural evolution across plant species?

Computational pipeline:
- Homology modeling: Use SWISS-MODEL with human APE1 (PDB: 4LND) as a template ( ).
- Docking simulations: Predict interactions with ROS1’s lyase domain using HADDOCK ( ).
- Conserved motif analysis: Align APE1L sequences from Arabidopsis, wheat (TaApe1L), and maize ( ).

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APE1L Antibody