Customize AKT5 Antibody

Description

Clarification of Terminology

AKT (Protein Kinase B): A serine/threonine kinase with three isoforms (AKT1, AKT2, AKT3) involved in cell survival, proliferation, and metabolism .
AK5 (Adenylate Kinase 5): A cytosolic enzyme regulating nucleotide metabolism and energy homeostasis, implicated in cancers, neurodegenerative diseases, and autoimmune encephalitis .

No "AKT5" gene or protein has been identified in peer-reviewed studies. The likely focus is AK5 antibodies, which are validated in research and clinical contexts.

Basic Characteristics

Target: AK5 (UniProt ID: Q9Y6K8), a 63 kDa protein .
Host/Isotype: Rabbit IgG (polyclonal) .
Tested Reactivity: Human, mouse, rat .

Applications

Application	Dilution Range	Detected Samples
Western Blot (WB)	1:1000–1:5000	Mouse brain, cerebellum; rat brain
Immunohistochemistry (IHC)	1:50–1:500	Mouse brain (antigen retrieval: TE/citrate buffer)
Immunofluorescence (IF)	1:50–1:500	Mouse brain

Clinical and Research Significance

Autoimmune Encephalitis: AK5-IgG antibodies are biomarkers for severe, non-paraneoplastic limbic encephalitis. Patients present with amnestic syndrome, mesiotemporal MRI hyperintensities, and hippocampal atrophy .
- Demographics: Predominantly affects elderly males (mean age ~70 years) .
- Diagnosis: Detected via tissue-based assays (TIF) and confirmed by cell-based assays (CBA) .
Cancer:
- Colorectal Cancer (CRC): AK5 promoter hypermethylation inversely correlates with expression, inhibiting metastasis via AMPK/mTOR pathway modulation .
- Breast/Gastric Cancer: AK5 regulates migration and invasion in cell lines .

Role in Disease Pathways

Neurodegeneration: AK5 dysfunction is linked to Alzheimer’s and Parkinson’s diseases through proteasome inhibition and energy metabolism disruption .
Autoimmunity: AK5 antibodies are associated with cytotoxic T-cell-mediated hippocampal damage, showing poor response to immunotherapy .

AK5 vs. AKT in Research

Feature	AK5	AKT (Pan)
Function	Nucleotide metabolism, energy balance	Cell survival, proliferation
Antibody Targets	Cytosolic enzyme	Phosphorylated (T308/S473) sites
Disease Links	Encephalitis, cancers	Diabetes, cancer, Proteus syndrome
Therapeutic Potential	Prognostic marker in CRC	Targeted therapies (e.g., MK-2206)

Limitations and Future Directions

Specificity Challenges: False positives reported in immunoblots/CBA .
Therapeutic Exploration: AK5’s role in AMPK/mTOR signaling warrants investigation for metastatic cancer interventions .

Product Specs

Buffer

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

Form

Liquid

Lead Time

14-16 week lead time (made-to-order)

Synonyms

AKT5 antibody; At4g32500 antibody; F8B4.200Probable potassium channel AKT5 antibody

Target Names

AKT5

Uniprot No.

Q9SCX5

Target Background

Function

This antibody targets a probable potassium channel protein. It may interact with the cytoskeleton or with associated regulatory proteins.

Database Links

KEGG: ath:AT4G32500

STRING: 3702.AT4G32500.1

UniGene: At.698

Protein Families

Potassium channel family, Plant (TC 1.A.1.4) subfamily

Subcellular Location

Membrane; Multi-pass membrane protein.

Tissue Specificity

Predominantly expressed in flowers.

Q&A

FAQs for Researchers on AKT5 Antibody

What experimental parameters critically influence AKT5 antibody performance in immunohistochemistry?

Parameter	Optimal Range	Impact on Results
Fixation time	18-24 hr (4% PFA)	Over-fixation masks epitopes
Antigen retrieval pH	6.0-8.0	pH <6 causes epitope denaturation
Antibody concentration	1:100-1:500	Higher concentrations increase background

How should researchers resolve contradictory AKT5 localization data between cytoplasmic and nuclear compartments?

Advanced analysis framework:
- Fractionation validation: Perform subcellular fractionation with compartment-specific markers (e.g., Lamin B1 for nuclear, GAPDH for cytoplasmic) .
- Phosphorylation-state analysis: Use phospho-specific antibodies to differentiate active/inactive forms (e.g., p-AKT5 vs total AKT5).
- Temporal profiling: Monitor localization changes across cell cycle stages using synchronized cultures .

What are robust negative controls for AKT5 functional studies in kinase assays?

Three-tier control system:
- Pharmacological: AKT inhibitor VIII (5µM) + antibody
- Genetic: siRNA-mediated AKT5 knockdown
- Isotype control: Non-immune IgG matched for host species

How to optimize AKT5 antibody cross-reactivity assessment across species?

Comparative immunoprecipitation protocol:
- Prepare lysates from human, mouse, and rat tissues
- Perform parallel IPs using same antibody lot
- Analyze precipitates via mass spectrometry
- Critical metric: ≥90% sequence homology in detected proteins

What statistical approaches are essential for AKT5 quantification in time-course experiments?

Advanced analytical pipeline:
- Mixed-effects modeling: Accounts for technical replicates and temporal autocorrelation
- Benjamini-Hochberg correction: Controls false discovery rate in high-throughput datasets
- Bootstrap validation: Assess stability of dose-response curves

How to troubleshoot AKT5 signal inconsistency between fresh and archived samples?

Diagnostic workflow:
- Epitope mapping: Confirm degradation-resistant epitopes using synthetic peptides
- Post-translational modification panel: Test for oxidation/phosphorylation changes in archived samples
- Antibody lot comparison: Include positive controls from original validation studies

What validation criteria distinguish research-grade from diagnostic-grade AKT5 antibodies?

Parameter	Research Grade	Diagnostic Grade
Lot-to-lot consistency	±15%	±5%
Stability (accelerated testing)	6 months	24 months
Cross-reactivity documentation	Major species	Full phylogenetic panel

Key Notes for Experimental Design:

Always include biological vs technical replicates matrix when testing AKT5 antibodies3.
For phospho-specific applications, maintain phosphatase inhibitors (e.g., 1mM Na3VO4) throughout processing .
In animal models, confirm antibody compatibility with strain-specific AKT5 isoforms before initiating long-term studies .

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AKT5 Antibody