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Description

Introduction to GK3 Antibody

The GK3 antibody targets glycerol kinase 3 (GK3), an enzyme encoded by the GK3P gene (UniProt ID: Q14409) that plays a critical role in glycerol metabolism and cellular energy regulation . This antibody is widely used in research to study GK3 expression and function across human, mouse, and rat models .

Immunogen and Reactivity

Immunogen: A synthetic peptide corresponding to residues 21–70 of human GK3 .
Reactivity: Validated for human, mouse, and rat tissues .
Specificity: Binds to both denatured (Western blot) and native (IHC) GK3 proteins .

Applications and Recommended Protocols

GK3 antibodies are optimized for multiple experimental workflows:

Application	Recommended Dilution	Validation Examples
Western Blot	1:500–1:2000	Detection in HeLa cells and 293 cell lysates
IHC	1:20–1:300	Staining in human skin cancer and gastric adenocarcinoma
ELISA	1:40,000	Quantitative analysis of GK3 levels

Research Findings and Validation Data

Functional Role: GK3 regulates glycerol uptake and metabolism, making it a biomarker for metabolic studies .
Diagnostic Utility: Strong immunoreactivity observed in brain tissues, suggesting relevance in neurological research .
Technical Validation:
- Western Blot: Clear bands at expected molecular weights in human cell lines .
- IHC: Robust staining in paraffin-embedded cancer tissues (e.g., gastric adenocarcinoma) .

Species Cross-Reactivity and Practical Considerations

Cross-Reactivity:
- Primate/Goat: Potential reactivity predicted via sequence homology but requires experimental validation .
- Negative Controls: No cross-reactivity reported in GPC3-negative cell lines (e.g., A431) .
Storage: Stable at -20°C for 12 months; avoid repeated freeze-thaw cycles .

Key Advantages and Limitations

Advantages	Limitations
High specificity across species	Unverified reactivity in non-mammalian systems
Broad application compatibility	Discrepancies in observed molecular weights
Validated for low-abundance targets	Limited clinical or therapeutic data available

Future Directions

Current research focuses on expanding GK3 antibody applications in metabolic disorder models and cancer biology. Collaborative studies are needed to explore therapeutic potential, particularly in diseases linked to glycerol metabolism dysregulation .

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

GK3 antibody; At3g06200 antibody; F28L1.14Guanylate kinase 3 antibody; chloroplastic antibody; AtGK3 antibody; EC 2.7.4.8 antibody; GMP kinase 3 antibody

Target Names

GK3

Uniprot No.

Q94JM2

Target Background

Function

GK3 Antibody is essential for recycling GMP and, indirectly, cGMP. It is also required for normal chloroplast development.

Database Links

KEGG: ath:AT3G06200

STRING: 3702.AT3G06200.1

UniGene: At.19098

Protein Families

Guanylate kinase family

Subcellular Location

Plastid, chloroplast.

Q&A

Here’s a structured FAQ collection for researchers working with GK3 antibodies, organized by complexity and grounded in experimental methodology:

Advanced Research Questions

How can cross-reactivity challenges with GK3 antibodies be resolved in multi-species studies?

Stepwise strategy:
- Pre-adsorb antibodies with recombinant GK3 homologs (human, mouse, rat) .
- Validate using species-specific cell lines (e.g., HeLa for human, NIH/3T3 for mouse) .
- Quantify off-target binding via surface plasmon resonance (SPR) with KD thresholds <10 nM .

What methodologies confirm GK3’s functional role in glycerol metabolism?

Functional assays:
- Enzyme activity: Measure glycerol kinase activity in GK3-overexpressing vs. knockout cells .
- Metabolic profiling: LC-MS quantification of glycerol-3-phosphate levels post-antibody blockade .
Pathway analysis: CRISPR interference coupled with RNA-seq to identify GK3-dependent metabolic networks.

How do researchers validate GK3 antibody specificity in CRISPR-modified models?

Validation Step	Experimental Design	Expected Outcome
CRISPR knockout	sgRNA targeting GK3 exon 2	Loss of WB band at 61 kDa
Rescue experiment	Transfect GK3 cDNA	Restoration of signal
Off-target screening	Whole-exome sequencing	No indels in homologous genes (e.g., GK1/GK2)

Data Contradiction Analysis

How to resolve discrepancies in GK3 detection across WB and ELISA?

Hypothesis testing:
- Post-translational modifications: Treat lysates with PNGase F to rule out glycosylation artifacts .
- Epitope masking: Compare linear vs. conformational epitopes using peptide-blocking assays .
Quantitative reconciliation: Normalize WB/ELISA data to GK3 activity assays (r² > 0.85 required) .

Emerging Techniques

Can generative AI improve de novo GK3 antibody design?

Case study: Anti-HER2 antibodies designed via deep learning achieved 10.6% binding rates for HCDR3 variants, with affinities rivaling trastuzumab (KD ~1 nM) .
Workflow adaptation:
- Train models on GK3 structural data (AlphaFold-predicted epitopes).
- Screen libraries via FACS-based ACE assays .

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GK3 Antibody