Customize GPAT7 Antibody

Description

Introduction to GPAT7 Antibody

The GPAT7 antibody is a specialized immunological tool designed to detect and study glycerol-3-phosphate acyltransferase 7 (GPAT7), a plant-specific enzyme critical for synthesizing suberin, a protective polyester in plant cell walls. This antibody enables researchers to investigate GPAT7's expression patterns, subcellular localization, and functional roles in stress responses and lipid metabolism .

Role in Suberin Biosynthesis

Overexpression Studies: Ectopic expression of GPAT7 in Arabidopsis results in accumulation of very-long-chain sn-1/3- and sn-2 monoacylglycerols (MAGs) and free fatty acids (C22:0, C24:0), mimicking GPAT5 activity .
Wound Response: GPAT7 expression is strongly induced by leaf wounding, correlating with suberin deposition at injury sites. gpat7 mutants fail to develop toluidine blue-staining resistance at wounds .

Table 2: Comparative Analysis of GPAT Isoforms

GPAT	Function	Substrate Preference	Phenotype of Knockout/Mutation
GPAT4	Cutin synthesis	C16:0, C18:1 ω-oxidized	Reduced cutin monomers
GPAT5	Suberin synthesis	Broad acyl-CoA range	Altered seed coat permeability
GPAT7	Suberin/wound response	Similar to GPAT5	Impaired wound healing

Evolutionary Conservation

GPAT7 homologs are conserved in land plants, with distinct clades emerging in bryophytes (GPAT4/6/8) and tracheophytes (GPAT5/7) . This evolutionary divergence aligns with specialized roles in cutin/suberin biosynthesis.

Applications of GPAT7 Antibody in Research

Localization Studies: Used to track GPAT7’s endoplasmic reticulum localization via immunofluorescence .
Expression Profiling: Quantifies GPAT7 upregulation under stress (e.g., wounding, pathogen attack) using Western blotting .
Genetic Validation: Confirms GPAT7 knockout/overexpression lines in model plants like Arabidopsis and Medicago sativa .

Technical Considerations

Cross-Reactivity: Antibody specificity must be validated against GPAT5 due to high sequence similarity .
Sample Preparation: Optimal detection requires microsomal fractionation to isolate membrane-bound GPAT7 .

Future Directions

Elucidate GPAT7’s interaction with cytochrome P450 oxidases in suberin monomer synthesis.
Engineer GPAT7-overexpressing crops for enhanced drought tolerance via suberin-mediated water retention .

Product Specs

Buffer

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

Form

Liquid

Lead Time

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

Synonyms

GPAT7; At5g06090; K16F4.5; Glycerol-3-phosphate acyltransferase 7; AtGPAT7

Target Names

GPAT7

Uniprot No.

Q9LHS7

Target Background

Function

This antibody targets GPAT7, an enzyme that catalyzes the esterification of an acyl-group from acyl-ACP to the sn-1 position of glycerol-3-phosphate. This is a critical step in glycerolipid biosynthesis.

Database Links

KEGG: ath:AT5G06090

STRING: 3702.AT5G06090.1

Protein Families

GPAT/DAPAT family

Subcellular Location

Membrane; Multi-pass membrane protein.

Tissue Specificity

Weakly or not expressed in roots, leaves, seedlings, developing siliques and flower buds.

Q&A

GPAT7 Antibody: Research-Grade FAQs

Advanced Research Questions

How to design experiments investigating GPAT7’s role in suberin biosynthesis?
- Step 1: Generate GPAT7-overexpressing Arabidopsis lines and analyze stem waxes for very-long-chain MAGs (C22:0, C24:0) via GC-MS .
- Step 2: Compare toluidine blue staining at wound sites in wild-type vs. gpat7 mutants to assess suberin deposition defects .
- Critical controls: Include GPAT5 mutants, as GPAT7 and GPAT5 share functional redundancy .
How to address contradictory GPAT7 localization data across studies?
- Issue: ER vs. mitochondrial localization claims.
- Resolution:
  - Use subcellular fractionation (e.g., differential centrifugation) followed by Western blotting with organelle-specific markers (Calnexin for ER, prohibitin for mitochondria) .
  - Overexpress FLAG-tagged GPAT7 and perform immunofluorescence colocalization assays .
What methods optimize GPAT7 antibody utility in lipidomic studies?
- Combine immunoprecipitation (IP) with LC-MS/MS to identify GPAT7-associated lipid species.
- Key parameters:
  - Use n-dodecyl-β-D-maltoside for membrane protein solubilization .
  - Validate IP efficiency via Western blot and activity assays (acyltransferase activity toward arachidonoyl-CoA) .

Table 1: GPAT7 Antibody Validation Parameters

Parameter	Specification	Source
Molecular Weight	57 kDa (calculated)
Immunogen	Recombinant fragment (aa 450–550)
Cross-reactivity	Human, Mouse (not tested in plants)
Critical Controls	siRNA knockdown, peptide blocking

Table 2: GPAT7 Functional Insights from Model Systems

System	Finding	Method
Arabidopsis	GPAT7 overexpression → C24:0 MAGs	GC-MS, GUS reporter assays
Human cell lines	ER localization confirmed	Subcellular fractionation, IF
ob/ob mice	No direct role in TAG synthesis	CD4+ T-cell proliferation assays

Methodological Notes

For lipidomics: GPAT7’s preference for arachidonoyl-CoA necessitates acyl-CoA substrate screens in vitro .
In plant studies: Wounding experiments (e.g., leaf mechanical injury) are critical to induce GPAT7 expression .
Antibody limitations: Does not distinguish between GPAT7 splice variants; validate via qRT-PCR alongside protein assays .

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GPAT7 Antibody