SYP51 Antibody

Definition and Target

SYP51 (Syntaxin of Plants 51) is a tonoplast-localized SNARE protein regulating vesicle fusion in plant endomembrane systems . The SYP51 antibody specifically recognizes this protein, enabling its visualization and functional analysis in cellular studies .

Production

• Antigen Design: Antibodies were generated using N-terminal fragments of SYP51 (amino acids 1–189) fused to a 6xHis tag, expressed in Escherichia coli strain BL21 (DE3) pLysS .

• Purification: Recombinant proteins were isolated via Ni-affinity chromatography and injected into rabbits for polyclonal antibody production .

• Specificity: Cross-reactivity tests confirmed no recognition of closely related syntaxins (e.g., SYP52, SYP61) .

Validation

Localization Studies

• TGN and PVC Association: Immunogold labeling in Arabidopsis roots revealed SYP51 colocalization with SYP21 (PVC marker) and SYP42 (TGN marker) .

• Tonoplast Dynamics: SYP51 antibodies identified tonoplast invaginations ("bulbing") during co-expression with ER aquaporin NIP1;1 .

Functional Insights

• Vacuolar Trafficking: SYP51 regulates the sorting of vacuolar markers (e.g., GFPgl133Chi) and interacts with SNARE complexes (VTI11, VAMP727) .

• Metal Transport: SYP51 knockout lines showed reduced copper uptake, linked to its role in membrane trafficking .

SNARE Complex Analysis

Proteomic studies of SYP61 compartments identified SYP51-associated proteins, including cellulose synthases and regulatory SNAREs, implicating it in exocytosis .

Interaction with NIP1;1

• Method: Ratiometric Bimolecular Fluorescence Complementation (rBiFC) confirmed SYP51-NIP1;1 interaction on the tonoplast .

• Outcome: This interaction modulates arsenic tolerance and vacuole morphology .

Limitations

• Degradation Artifacts: RFP-tagged SYP51 overexpressed in plants showed partial proteolysis, necessitating careful immunoblot interpretation .

• Cell-Type Specificity: Localization varies between root and leaf tissues, requiring context-specific validation .

Colocalization Frequency of SYP51 with Organelle Markers