PLD6 Antibody, FITC conjugated
Description
Introduction to PLD6 Antibody, FITC Conjugated
PLD6 (phospholipase D family member 6) is a mitochondrial cardiolipin hydrolase critical for mitochondrial fusion, piRNA biogenesis, and transposon repression during spermatogenesis . PLD6 Antibody, FITC conjugated refers to a fluorescently labeled polyclonal or monoclonal antibody designed for specific detection of PLD6 in research applications such as immunofluorescence (IF), flow cytometry (FCM), and Western blot (WB) . The FITC (fluorescein isothiocyanate) conjugation enables visualization under fluorescence microscopy or flow cytometers with excitation/emission maxima at 499/515 nm .
Epitope and Cross-Reactivity:
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Targets internal regions of PLD6, ensuring specificity against conserved domains .
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Cross-reactivity with mouse and rat orthologs is reported but requires validation .
Primary Uses:
Optimization Tips:
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Fixation: Use 4% paraformaldehyde for intracellular staining .
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Antigen Retrieval: EDTA buffer (pH 8.0) enhances epitope exposure .
Western Blot Analysis:
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Sample: Human THP-1 whole-cell lysates.
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Result: A single band at ~28 kDa, aligning with PLD6’s predicted molecular weight .
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Protocol: SDS-PAGE (5–20% gel), transferred to nitrocellulose membrane, and detected via ECL .
Immunohistochemistry:
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Tissue: Human endometrial adenocarcinoma.
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Observation: Strong cytoplasmic staining in cancer cells, validated using DAB chromogen .
Flow Cytometry:
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Cell Line: MCF-7 (human breast cancer).
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Signal: Distinct fluorescence shift compared to isotype controls, confirming specificity .
Supplier Landscape and Product Comparison
Research Findings Using PLD6 Antibodies
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Mitochondrial Fusion Mechanism: PLD6-generated phosphatidic acid (PA) recruits NME3 to mitochondrial contact sites, facilitating membrane tethering . Depleting NME3 disrupts PLD6-induced mitochondrial clustering .
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piRNA Biogenesis: PLD6-deficient mice exhibit retrotransposon activation due to defective piRNA processing, highlighting its role in genome stability .
Limitations and Considerations
Product Specs
Target Background
This antibody targets PLD6 (Phospholipase D6), also known as MitoPLD, an enzyme exhibiting both phospholipase and nuclease activities depending on cellular context. Its interaction with Mitoguardin (MIGA1 or MIGA2) influences dimer conformation, modulating the balance between lipase and nuclease activities. PLD6 plays a crucial role in mitochondrial dynamics, specifically fusion and fission, through its phospholipase activity. As a phospholipase, it utilizes cardiolipin, a mitochondrial lipid, as a substrate to produce phosphatidic acid (PA), a lipid second messenger. PA generation promotes Mitofusin-mediated mitochondrial fusion, while its subsequent cleavage by Lipin phosphatases yields diacylglycerol (DAG), favoring mitochondrial fission. Both Lipin and DAG regulate mitochondrial dynamics, influencing fusion and fission processes crucial for adapting mitochondrial metabolism to cellular needs. Mitochondrial fusion is particularly important for meeting increased nucleotide demands during DNA synthesis. Given the close relationship between mitochondrial function and dynamics, PLD6's activity is linked to cell growth, proliferation, and differentiation. Furthermore, PLD6 mediates MYC activity, promotes mitochondrial fusion, and activates AMPK, which subsequently inhibits YAP/TAZ, thereby stimulating cell growth and proliferation. PLD6's endonuclease activity is essential for PIWI-interacting RNA (piRNA) biogenesis during spermatogenesis, contributing to sperm fertility. Its single-strand-specific nuclease activity is critical for piRNA biogenesis/maturation. MOV10L1, which interacts with PLD6, facilitates the delivery of piRNA precursors to the enzyme for initial cleavage, generating piRNA intermediate fragments that subsequently bind to Piwi proteins. PLD6 cleaves both DNA and RNA substrates with similar efficiency, producing a 5' phosphate end, participating in the processing of primary piRNA transcripts. piRNAs are vital for protecting against the activity of mobile genetic elements; piRNA-mediated transposon silencing maintains genome stability, especially in germline cells where transposon mobilization is prevalent due to widespread genomic demethylation. PA may function as a signaling molecule in the recognition and transport of primary piRNA precursor RNAs. Finally, PLD6 interaction with tesmin in testes suggests a role in spermatogenesis through this interaction.
- MIGA proteins promote mitochondrial fusion by regulating mitochondrial phospholipid metabolism via MitoPLD. PMID: 26711011
- MitoPLD plays a critical role in a pathway generating a specialized form of RNAi necessary for suppressing transposon mobilization during meiosis in developing spermatocytes. PMID: 22281006
- Mitochondrial-surface phosphatidic acid, generated by MitoPLD/Zuc, recruits or activates nuage components essential for piRNA production. PMID: 21397848
- PLD6 (MitoPLD) contains an N-terminal mitochondrial localization sequence, resides on the outer mitochondrial membrane, generates phosphatidic acid (a signaling lipid), and triggers trans-mitochondrial interactions; implicated in mitochondrial fusion. PMID: 17028579
