pept-1 Antibody
Description
Applications in Research
PEPT1 antibodies are widely used in:
Key Findings from Antibody Validation Studies:
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Species Reactivity: Confirmed in human, mouse, rat, cow, and chicken .
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Knockout Validation: No cross-reactivity observed in Pept1 −/− mice .
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Tissue Specificity:
Role in Therapeutic Research
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Cancer Targeting:
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Inflammatory Bowel Disease:
Research Implications
PEPT1 antibodies have clarified the transporter’s role in:
Product Specs
Target Background
PMID: 24999909, Intestinal amino acid availability, mediated by PEPT-1, affects TORC1/2 signaling and the unfolded protein response., .
PMID: 19621081, PEPT-1 loss decreases intestinal proton influx, resulting in increased free fatty acid uptake and fat accumulation., .
PMID: 15155758, PEPT-2 interacts with both the *C. elegans* target of rapamycin (TOR) and the DAF-2/insulin-signaling pathways., .
Q&A
Basic Research Questions
How to validate PEPT1 antibody specificity in immunohistochemistry (IHC)?
Methodological Answer:
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Pre-absorption controls: Pre-incubate the antibody with excess PEPT1 peptide antigen to confirm reduced/no binding .
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Cell line validation: Compare PEPT1 expression in positive controls (e.g., Caco-2 cells) vs. negative controls (e.g., NCM460 cells) using RT-qPCR and Western blot .
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Tissue specificity: Use tissue microarrays to compare PEPT1 expression in cancer vs. adjacent normal tissues .
Table 1: Key Validation Metrics for PEPT1 Antibodies
What are the most reliable experimental models for studying PEPT1 in cancer?
Methodological Answer:
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In vitro: Use hepatocarcinoma (HepG2) or pancreatic cancer cell lines (e.g., PDAC) with confirmed PEPT1 overexpression via RT-qPCR .
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In vivo: Employ xenograft models (e.g., mouse pancreatic cancer xenografts) with shRNA knockdown to assess PEPT1’s role in tumor growth .
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Tumor microenvironment: Mimic lactic acid-rich conditions (Warburg effect) to study PEPT1’s transport activity .
Table 2: Model Systems for PEPT1 Studies
Advanced Research Questions
How to resolve contradictory data on PEPT1 expression across cancer types?
Methodological Answer:
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Pathological grading: Stratify samples by tumor grade (e.g., PEPT1 expression increases with malignancy in hepatocarcinoma) .
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Microenvironmental factors: Account for lactic acid levels, which upregulate PEPT1 transport activity in pancreatic cancer .
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Antibody variability: Compare multiple clones (e.g., clone 119 vs. 253) with distinct inhibition efficiencies .
Table 3: Contradictory Findings in PEPT1 Literature
What functional assays confirm PEPT1 antibody efficacy in transport inhibition?
Methodological Answer:
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Radiolabeled substrate competition: Use [³H]-glycylsarcosine uptake assays with/without anti-PEPT1 antibodies (e.g., clone 119 inhibits 20% activity) .
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Pharmacological inhibition: Combine antibodies with DPPIV/MMP inhibitors to block peptide substrate generation .
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In vivo targeting: Measure NP accumulation in colitis models via PepT1-mediated uptake (e.g., FK506-loaded NPs reduce TNF-α by 50%) .
Table 4: Functional Assays for PEPT1 Antibodies
How to design PEPT1-targeted drug delivery systems for in vivo applications?
Methodological Answer:
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Nanoparticle (NP) functionalization: Use co-assembly of anti-inflammatory peptides (e.g., KPV) and immunosuppressants (e.g., FK506) with PepT1-binding ligands .
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Biodistribution analysis: Track NPs in DSS-induced colitis models via immunofluorescence (e.g., colocalization with PepT1 in inflamed epithelia) .
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Dose optimization: Balance peptide ligand density (≥0.48 propensity score) to maximize targeting and minimize off-site effects .
