FER1 Antibody
Description
Definition and Molecular Targets
The term "FER1" refers to different proteins depending on the organism:
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Chlamydomonas FER1: A ferritin subunit involved in iron storage and homeostasis. It forms part of a 25 kDa monomeric protein complex, distinct from the trimeric FER2 ferritin ( ).
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Human/mouse FER (FER1): A proto-oncogene-encoded tyrosine kinase (UniProt: P16591) regulating cell adhesion, migration, and immune responses. Commercial FER1 antibodies (e.g., Proteintech 25287-1-AP) target this protein ( ).
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Toxoplasma FER1: A ferlin-family protein (159 kDa) critical for microneme organelle trafficking and exocytosis in Toxoplasma gondii ( ).
Research-Grade Antibodies
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Toxoplasma FER1: Polyclonal antiserum against the C2DE domain detects full-length FER1 (159 kDa) and fragments (120 kDa, 30 kDa) in Western blots ( ).
Ferritin Studies (Chlamydomonas)
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Anti-FER1 antibodies immunopurify ferritin1 complexes, revealing a 70:1 abundance ratio of FER1:FER2 subunits under iron-replete conditions ( ).
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Subcellular localization studies demonstrate FER1’s role in cytoplasmic iron sequestration ( ).
Cancer and Cell Biology (Human/Mouse)
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Proteintech’s FER1 antibody detects FER tyrosine kinase in:
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Overexpression of FER correlates with tumor progression and chemoresistance in colorectal cancer models ( ).
Parasite Biology (Toxoplasma)
Technical Considerations
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
- Research indicates that the phosphate starvation response of AtFer1 is not directly linked to the iron status of plants but is specifically triggered by phosphate deficiency. PMID: 23788639
- Studies have shown that a leaf glutathione concentration threshold between 10 and 50 nmol GSHg(-1) FW is required for the full induction of AtFer1 gene expression in response to iron. PMID: 22449975
- Investigations have revealed a novel regulatory pathway involved in plant response to oxidative stress using the iron-induced Arabidopsis ferritin AtFER1 as a model system. PMID: 21946558
- Ferritin accumulation during the infection of Arabidopsis by E. chrysanthemi is a fundamental defense mechanism primarily activated by bacterial siderophores. PMID: 15998312
Q&A
Basic Research Questions
How to select the appropriate FER1 antibody for different experimental designs?
Choose antibodies based on three factors:
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Application specificity: Western blot (WB) requires antibodies recognizing denatured epitopes (e.g., Proteintech 67671-1-PBS targets FER in WB/IF/ICC ), while immunofluorescence (IF) needs antibodies against native conformations.
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Epitope region: For functional studies, select antibodies targeting domains critical for FER1 activity (e.g., C2DE domain antibodies in Toxoplasma FER1 studies ).
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Validation rigor: Prioritize antibodies with knockout-validated specificity (e.g., fer-1 mutant controls in C. elegans ).
| Antibody Performance Across Studies |
|---|
| Application |
| ------------------ |
| WB/IF/ICC |
| ImmunoEM |
| Subcellular IFA |
What methods validate FER1 antibody specificity in complex biological systems?
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Western blot: Compare observed (95 kDa for human FER ) vs. predicted molecular weight.
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Knockout controls: Use FER1 deletion mutants (e.g., Chlamydomonas FER1/2 iron-response studies ).
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Orthogonal localization: Confirm subcellular patterns with independent markers (e.g., FER1 apical-nuclear localization vs. MIC2 in Toxoplasma ).
How to troubleshoot nonspecific bands in FER1 antibody-based assays?
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Test multiple antibodies against distinct domains (e.g., C2DE vs. DysfNC domains ).
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Pre-absorb antibodies with recombinant FER1 protein (see C. elegans immunoelectron microscopy protocols ).
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Validate using conditional depletion models (e.g., Shield-1-stabilized dominant-negative FER1 mutants ).
Advanced Research Questions
How to resolve contradictory FER1 localization data across studies?
Contradictions arise from:
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Environmental stimuli: FER1 relocalizes from apical-nuclear regions (intracellular parasites) to cytoplasm (extracellular) in Toxoplasma .
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Fixation artifacts: Use live-cell imaging tags (e.g., DD-Myc-FER1∆TM ) to bypass antibody-dependent methods.
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Species-specific isoforms: Compare Chlamydomonas FER1 (chloroplastic ) vs. human FER (cytosolic ).
| Localization Discrepancies |
|---|
| System |
| --------------------- |
| Toxoplasma |
| Toxoplasma |
| C. elegans sperm |
How to design experiments analyzing FER1's role in calcium-dependent exocytosis?
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Functional assays: Combine propranolol-induced Ca²⁺-independent secretion with FER1 antibody-based microneme tracking .
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Genetic perturbations: Use conditional FER1 mutants (e.g., auxin-inducible degron) alongside quantitative IF .
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Dynamic imaging: Employ time-lapse microscopy with FER1/MIC2 dual labeling to monitor trafficking .
What strategies address cross-reactivity in cross-species FER1 studies?
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Epitope conservation analysis: Align sequences across species (e.g., human FER vs. Chlamydomonas FER1 ).
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Empirical validation: Test antibody reactivity in phylogenetically distant systems (e.g., C. elegans vs. mammalian cells ).
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Domain-specific blocking: Pre-incubate antibodies with recombinant proteins from non-target species.
How to interpret FER1 antibody data in systems with paralogs (e.g., FER1 vs. FER2)?
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Subunit-specific antibodies: Use antibodies targeting non-conserved regions (e.g., Chlamydomonas FER1 vs. FER2 ).
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Quantitative proteomics: Combine immunoprecipitation with mass spectrometry to distinguish paralog contributions.
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Gene silencing: Perform RNAi knockdown of individual paralogs followed by antibody validation .
