PUMP1 Antibody
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
- Transcriptomic analysis of overexpressing plants revealed a broad induction of stress-responsive genes not strictly related to the mitochondrial antioxidant machinery. This suggests that overexpression of AtUCP1 induces a strong stress response within the cell. PMID: 26494730
- Overexpression of UCP1 in the mitochondrial inner membrane resulted in increased uncoupled respiration, decreased reactive oxygen species accumulation under abiotic stresses, and diminished cellular ATP content. PMID: 24886177
- The primary physiological role of UCP1 in Arabidopsis leaves is related to maintaining the redox poise of the mitochondrial electron transport chain to facilitate photosynthetic metabolism. PMID: 17148605
- The Cys28 substitution reduced the ATP inhibitory effect, while the Tyr269Phe mutant exhibited a 2.8-fold increase in sensitivity to ATP. PMID: 17980348
Q&A
Here’s a structured FAQ collection for researchers working with PUMP1 antibodies, organized by scientific depth and methodological rigor:
How do I validate the specificity of PUMP1 antibodies in immunoassays?
Methodology:
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Perform western blotting with positive/negative controls (e.g., cell lysates expressing PUMP1 vs. knockout models) .
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Use blocking peptides to confirm antibody-antigen binding specificity .
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Validate via immunoprecipitation followed by mass spectrometry to verify target identity.
Key Controls:
| Control Type | Purpose | Example |
|---|---|---|
| Isotype Control | Non-specific binding baseline | IgG from same host species |
| Knockout Validation | Confirm absence of signal in PUMP1-deficient samples | CRISPR-edited cell line |
| Cross-Reactivity Check | Assess off-target binding | Lysates from tissues with homologous proteins |
What protocols optimize PUMP1 antibody performance in flow cytometry?
Methodology:
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Cell Preparation: Maintain >90% viability to reduce background noise; use PBS with 0.1% sodium azide to prevent antigen internalization .
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Titration: Test antibody concentrations (e.g., 0.1–2 µg/mL) to determine optimal signal-to-noise ratio.
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Compensation: Use single-stained controls for spectral overlap correction .
Critical Parameters:
How can Design of Experiments (DOE) optimize PUMP1 antibody-based assay conditions?
Methodology:
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Apply full factorial designs to evaluate interactions between variables (e.g., pH, incubation time, temperature) .
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Define responses (e.g., signal intensity, non-specific binding) and use software (e.g., MODDE®) to model Design Space .
Example DOE Setup:
| Factor | Low Level | High Level |
|---|---|---|
| pH | 6.8 | 7.4 |
| Incubation Time | 30 min | 60 min |
| Antibody Concentration | 0.5 µg/mL | 2 µg/mL |
Outcome: Identifies robust conditions minimizing variability during scale-up .
How should researchers resolve contradictory data from PUMP1 antibody studies?
Methodology:
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Variable Audit: Compare protocols for differences in fixation methods, buffer composition, or epitope accessibility .
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Cross-Platform Validation: Replicate findings using orthogonal methods (e.g., immunohistochemistry vs. ELISA) .
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Reagent Traceability: Document antibody lot numbers and storage conditions to rule out batch effects .
Case Example:
Discrepancies in subcellular localization may arise from fixation artifacts. Validate using multiple fixation protocols (e.g., paraformaldehyde vs. methanol) .
What strategies enhance reproducibility of PUMP1 antibody studies in disease models?
Methodology:
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Tissue-Specific Validation: Test antibody performance in disease-relevant tissues (e.g., tumor vs. healthy biopsies) .
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Multiplexed Assays: Combine PUMP1 detection with biomarkers (e.g., MMP-7, MMP-12) to contextualize findings .
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Blinded Analysis: Minimize bias by encoding samples during data acquisition/interpretation .
Data Integration:
| Biomarker Panel | Purpose |
|---|---|
| PUMP1 + MMP-12 | Assess metastatic potential in lung cancer models |
| PUMP1 + HLA Class I | Investigate immune evasion mechanisms |
