PTCH2 Antibody, FITC conjugated
Description
Introduction
The PTCH2 Antibody, FITC conjugated is a fluorescently labeled immunological reagent designed to detect and study the Patched 2 (PTCH2) protein, a critical receptor in the Hedgehog signaling pathway. This antibody combines a polyclonal or monoclonal anti-PTCH2 antibody with fluorescein isothiocyanate (FITC), enabling visualization in fluorescence-based assays such as immunofluorescence microscopy, flow cytometry, and fluorescent Western blotting .
Core Components
Key Functional Properties
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Specificity: Recognizes PTCH2 in human tissues, particularly in skin and spermatocytes .
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Affinity: FITC labeling may reduce binding affinity compared to unconjugated antibodies, necessitating careful optimization .
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Applications: Primarily used for detecting PTCH2 localization in fixed or live cells, though exact applications may require user validation .
Fluorescence-Based Detection
Critical Insights
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Hedgehog Signaling: PTCH2 forms a receptor complex with Smoothened (SMO) and binds Hedgehog ligands (e.g., Sonic Hedgehog, Shh) . FITC-conjugated antibodies enable real-time tracking of PTCH2-SMO interactions in live-cell imaging.
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Non-Canonical Pathways: PTCH2 mediates cAMP/Creb and Src kinase signaling independently of Gli transcription factors, as shown in germ cell studies .
PTCH2’s Role in Hedgehog Signaling
Key Implications
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Tissue-Specific Signaling: PTCH2/Gas1 complexes in germ cells and PTCH1/Boc complexes in somatic cells suggest divergent roles in Hedgehog signal transduction .
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Disease Relevance: Altered PTCH2 expression is linked to basal cell carcinoma and medulloblastoma, making it a therapeutic target .
Example Product Profile (ABIN7165909)
| Parameter | Value | Notes |
|---|---|---|
| Target | PTCH2 (AA 793–951) | Human-specific epitope |
| Host | Rabbit | Polyclonal antibody |
| Conjugate | FITC | Requires light protection |
| Reconstitution | 100 μg/vial (lyophilized) | Reconstitute in PBS or distilled water |
| Storage | -20°C (long-term), 2–8°C (short-term) | Avoid repeated freeze-thaw cycles |
Optimal FITC Conjugation Parameters
| Factor | Optimal Condition | Impact | Source |
|---|---|---|---|
| pH | 9.5 | Maximizes FITC-protein coupling | |
| Temperature | Room temperature (25°C) | Rapid reaction completion | |
| Protein Concentration | 25 mg/mL | High labeling efficiency |
Challenges and Recommendations
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Labeling Index: Higher FITC labeling reduces binding affinity but improves sensitivity. Optimize dilution (e.g., 0.1–0.5 μg/mL for Western blot) .
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Cross-Reactivity: Validate specificity using blocking peptides or isotype controls .
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Assay Design: Pair with anti-FITC secondary antibodies for signal amplification in flow cytometry .
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
- Combined heterozygous germline mutations in PTCH1 and PTCH2 have been identified in a patient with embryonal rhabdomyosarcoma. PMID: 29230040
- Frameshift mutations in the PTCH2 gene can lead to nevoid basal cell carcinoma syndrome. PMID: 23479190
- PTCH2 isoforms exhibit distinct roles in Hedgehog signaling. PMID: 14613484
- The novel missense mutation PTCH2 (2157G-->A) underlies NBCCS, resulting in the loss of PTCH2's inhibitory function in the Shh signaling pathway. PMID: 18285427
- A susceptibility locus on 1p32-1p34 for congenital macrostomia in a Chinese family and the identification of a novel PTCH2 mutation have been reported. PMID: 19208383
