MitHPPK/DHPS Antibody
Description
Antibody Validation Data
Commercial antibodies validated for DHPS detection show cross-reactivity with mitHPPK/DHPS due to conserved epitopes:
Key disparities in observed molecular weights (35–72 kDa) suggest isoform-specific detection or post-translational modifications.
Folate Biosynthesis Studies
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Structural analyses using mitHPPK/DHPS antibodies revealed its role in herbicidal targeting, with sulfonamide inhibitors showing cross-kingdom efficacy against plant and microbial forms .
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In Arabidopsis, mitHPPK/DHPS knockout is lethal, while cytosolic forms regulate stress responses .
Inflammation and Metabolism
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DHPS hypusinates eIF5A, promoting proinflammatory M1 macrophage polarization. Antibodies enabled quantification of DHPS-eIF5A complexes in obese adipose tissue, linking enzyme activity to glucose intolerance .
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Myeloid-specific Dhps knockout mice showed reduced adipose M1 macrophages and improved insulin sensitivity .
Technical Considerations
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Dilution optimization: WB concentrations range from 1:200 (polyclonal ) to 1:50,000 (monoclonal ).
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Sample preparation: Mitochondrial fractionation is critical for mitHPPK/DHPS detection, as cytosolic isoforms may cross-react .
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Validation controls: Peptide blocking experiments (e.g., Boster Bio’s A30637 ) confirm antibody specificity.
Limitations and Future Directions
Current antibodies lack explicit validation for mitochondrial-specific HPPK/DHPS. Structural modeling data suggest epitopes in the linker region (residues 161–202) could be targeted to develop isoform-selective antibodies. Proteomic studies using existing reagents have identified DHPS interactions with NF-κB pathway components, highlighting its therapeutic potential in metabolic diseases .
Product Specs
Composition: 50% Glycerol, 0.01M Phosphate Buffered Saline (PBS), pH 7.4
Target Background
Q&A
Here’s a structured FAQ guide for researchers working with DHPS (Deoxyhypusine Synthase) antibodies, based on academic research scenarios and curated from the provided sources:
Advanced Research Questions
How to resolve discrepancies in DHPS banding patterns across studies?
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Approach:
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Post-translational modifications: DHPS undergoes hypusination, altering electrophoretic mobility. Use enzymatic deglycosylation or phosphatase treatment .
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Antibody clonality: Polyclonal antibodies (e.g., ab190266) may detect multiple isoforms, while monoclonal (e.g., 67998-1-Ig) show higher specificity .
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Buffer optimization: Adjust SDS-PAGE conditions (e.g., 10–12% gels) to improve resolution of ~40 kDa bands .
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What strategies enhance DHPS detection in low-abundance samples?
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Signal amplification:
How to validate cross-species reactivity for DHPS antibodies?
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Stepwise validation:
