PTCH2 Antibody
Description
Introduction to PTCH2 Antibody
PTCH2 Antibody refers to specialized immunological reagents developed to detect and bind to Protein patched homolog 2 (PTCH2). These antibodies serve as critical tools for scientific investigation, allowing researchers to examine PTCH2 expression, localization, and function across different tissues and cell types. PTCH2 is a transmembrane protein that plays significant roles in multiple cellular processes, particularly within the Sonic Hedgehog (SHH) signaling pathway, making reliable detection methods essential for advancing research in developmental biology and cellular growth regulation .
Multiple manufacturers produce PTCH2 antibodies, including Cell Signaling Technology, Bio-Techne/R&D Systems, Abcam, and Boster, each offering products with specific characteristics and applications . These antibodies are available in various forms, including monoclonal and polyclonal variants, and are designed to react with PTCH2 from different species, most commonly human and mouse specimens.
Structure and Function of PTCH2 Protein
Understanding the target protein's structure provides important context for antibody applications. PTCH2 (Protein patched homolog 2) is a multi-pass transmembrane protein with a molecular weight of approximately 130-180 kDa . The mouse PTCH2 protein consists of 1182 amino acids and contains two terminal cytoplasmic domains between amino acids 1-57 and 1108-1182 . The protein features 12 transmembrane domains that are critical to its function.
PTCH2, together with Smoothened (SMO), forms a membrane complex that serves as a receptor for Sonic Hedgehog (SHH) . This complex plays a crucial role in the Hedgehog signaling pathway, which regulates various developmental processes. PTCH2 demonstrates less widespread expression than its homolog PTCH1 but shows particularly high expression in skin tissues and spermatocytes .
The primary biological functions of PTCH2 include:
Monoclonal PTCH2 Antibodies
Monoclonal PTCH2 antibodies derive from single B-cell clones, resulting in highly specific antibodies targeting particular epitopes of the PTCH2 protein. Several commercial monoclonal PTCH2 antibodies are available:
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Rabbit Monoclonal Anti-PTCH2 [EPR26073-39] from Abcam (ab300036, ab300037): This antibody demonstrates suitability for Western Blotting, Immunohistochemistry, Immunocytochemistry/Immunofluorescence, Flow Cytometry, and Immunoprecipitation applications with human samples .
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Mouse Monoclonal Anti-Human Patched 2/PTCH2 (Bio-Techne, MAB4078, Clone # 403438): This antibody works effectively for Immunohistochemistry and Western Blot applications for detecting human PTCH2 .
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Rat Monoclonal Anti-Mouse Patched 2/PTCH2 (R&D Systems, MAB8096, Clone # 865619): Specifically designed for detecting mouse PTCH2 in immunofluorescence applications .
Monoclonal antibodies offer significant advantages including consistent performance between production batches and high specificity for target epitopes, making them valuable for precise detection of PTCH2 in research applications.
Polyclonal PTCH2 Antibodies
Polyclonal PTCH2 antibodies derive from multiple B-cell clones and recognize multiple epitopes on the PTCH2 protein. Notable examples include:
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Rabbit Polyclonal Anti-PTCH2 from Boster Bio (PA1859): This antibody functions primarily in Western Blot applications and reacts with both human and mouse PTCH2 .
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Rabbit Polyclonal Anti-PTCH2 (L849) and Anti-PTCH2 (G1191) from Cell Signaling Technology (#2464 and #2470): These antibodies detect transfected levels of PTCH2 protein and demonstrate no cross-reactivity with human PTCH1 .
Polyclonal antibodies typically offer enhanced sensitivity for detection since they bind to multiple epitopes on the target protein simultaneously, potentially increasing signal strength in applications with low abundance targets.
Applications of PTCH2 Antibodies
PTCH2 antibodies serve multiple laboratory techniques designed to investigate expression, localization, and function of the PTCH2 protein. The major applications include:
Western Blotting Applications
Western Blotting represents a fundamental technique for detecting specific proteins in biological samples. PTCH2 antibodies are widely employed in Western Blotting to detect PTCH2 protein expression across various tissues and cell lines .
For example, Mouse Anti-Human Patched 2/PTCH2 Monoclonal Antibody (MAB4078) has successfully detected PTCH2 in lysates from OVCAR-3 human ovarian carcinoma cells, WS-1 human fetal skin fibroblasts, and Bowes human melanoma cell lines . This antibody detected a specific band for PTCH2 at approximately 280 kDa under reducing conditions.
| Antibody | Recommended Dilution | Source | Applications |
|---|---|---|---|
| PTCH2 (L849) #2464 | 1:1000 | Rabbit | WB, IP |
| PTCH2 (G1191) #2470 | 1:1000 | Rabbit | WB, IP |
| Anti-PTCH2 PA1859 | Not specified | Rabbit | WB |
| MAB4078 | Not specified | Mouse | WB, IHC |
Immunoprecipitation Studies
Immunoprecipitation techniques isolate specific proteins from complex mixtures using antibody-based capture. Several PTCH2 antibodies demonstrate suitability for immunoprecipitation applications .
The Rabbit Monoclonal Anti-PTCH2 [EPR26073-39] antibody (ab300036) has successfully immunoprecipitated PTCH2 from SH-SY5Y human neuroblastoma epithelial cell whole cell lysate . For optimal results, manufacturers typically recommend a 1:50 dilution for PTCH2 antibodies in immunoprecipitation applications .
Immunohistochemistry Applications
Immunohistochemistry techniques detect specific proteins within tissue sections. Multiple PTCH2 antibodies demonstrate efficacy for immunohistochemical applications .
The Mouse Anti-Human Patched 2/PTCH2 Monoclonal Antibody (MAB4078) has successfully detected PTCH2 in immersion-fixed paraffin-embedded sections of human cervical cancer tissue at a concentration of 15 µg/mL . The analysis revealed specific staining localized to cytoplasm in epithelial cells, providing valuable information about PTCH2 distribution within these tissues.
Similarly, Rat Anti-Mouse Patched 2/PTCH2 Monoclonal Antibody (MAB8096) effectively detected PTCH2 in immersion-fixed frozen sections of mouse testis at a concentration of 5 µg/mL , demonstrating the versatility of these antibodies across different species and tissue preparation methods.
Flow Cytometry Applications
Flow cytometry analyzes characteristics of individual cells within heterogeneous populations. Select PTCH2 antibodies, such as Rabbit Monoclonal Anti-PTCH2 [EPR26073-39] (ab300036), demonstrate compatibility with intracellular flow cytometry applications , providing researchers with tools for quantitative analysis of PTCH2 expression at the cellular level.
Species Reactivity Profiles
PTCH2 antibodies demonstrate species-specific recognition patterns that researchers must consider when selecting appropriate reagents. The predominant reactivity profiles include:
Human-Reactive PTCH2 Antibodies
Numerous commercially available antibodies target human PTCH2, including products from Abcam (ab300036, ab300037), Bio-Techne (MAB4078), and Cell Signaling Technology (#2464, #2470) . These antibodies enable investigation of PTCH2 in human tissues, cell lines, and clinical samples.
Mouse-Reactive PTCH2 Antibodies
For research employing mouse models, antibodies such as Rat Monoclonal Anti-Mouse Patched 2/PTCH2 (MAB8096) from R&D Systems and Rabbit Polyclonal Anti-PTCH2 (PA1859) from Boster Bio provide specific detection of mouse PTCH2 .
Some antibodies, including Rabbit Polyclonal Anti-PTCH2 (PA1859) from Boster Bio, demonstrate cross-reactivity with both human and mouse PTCH2 , offering versatility for comparative studies across species.
PTCH2 in Growth Regulation
Recent research highlights PTCH2's involvement in cellular growth control mechanisms . The protein may influence proliferation pathways, making PTCH2 antibodies valuable tools for studying developmental processes and growth regulation in both normal and pathological contexts.
PTCH2 in Sonic Hedgehog Signaling
PTCH2 forms a critical receptor complex with Smoothened (SMO) that responds to Sonic Hedgehog (SHH) ligands . This signaling pathway plays fundamental roles in embryonic development, tissue patterning, and stem cell maintenance. PTCH2 antibodies enable researchers to visualize and quantify PTCH2 expression and distribution within this important signaling pathway.
PTCH2 in Mesenchymal Stem Cell Regulation
A 2022 study provided evidence that PTCH2 may function as a regulator of mesenchymal stem cells (MSCs) . Researchers observed significant changes in incisor mesenchymal stem cell niches that contributed to accelerated incisor growth in Ptch2 knockout mice (Ptch2−/−). The study noted changes specifically in tissues where Ptch2 co-expresses with Ptch1, affecting MSC regenerative and differentiation potential .
Importantly, absence of PTCH2 correlated with increased stem cell functionality, suggesting PTCH2 plays an independent role in regulating stem cell function and postnatal regenerative capacity . This research highlights the value of PTCH2 antibodies for investigating stem cell biology and tissue regeneration mechanisms.
Table 1: PTCH2 (L849) Antibody #2464 - Cell Signaling Technology
| Parameter | Specification |
|---|---|
| Reactivity | Human |
| Sensitivity | Transfected Only |
| Molecular Weight | 130 kDa |
| Source | Rabbit |
| Applications | Western Blotting, Immunoprecipitation |
| Dilution for Western Blotting | 1:1000 |
| Dilution for Immunoprecipitation | 1:50 |
| Cross-Reactivity | Does not recognize human PTCH1 |
Table 2: Human Patched 2/PTCH2 Antibody (MAB4078) - Bio-Techne
| Parameter | Specification |
|---|---|
| Species Reactivity | Human |
| Applications | Immunohistochemistry, Western Blot |
| Label | Unconjugated |
| Antibody Source | Monoclonal Mouse IgG |
| Clone | 403438 |
| Immunogen | E. coli-derived recombinant human Patched 2/PTCH2 (Glu79-Glu392) |
| Accession | Q9Y6C5 |
| Specificity | Detects human Patched 2/PTCH2 in direct ELISA and Western Blots |
| Recommended Usage for IHC | 8-25 µg/mL |
Table 3: Mouse Patched 2/PTCH2 Antibody (MAB8096) - R&D Systems
| Parameter | Specification |
|---|---|
| Clone | 865619 |
| Immunogen | E. coli-derived recombinant mouse Patched 2/PTCH2 (Glu79-Ser394) |
| Accession | O35595 |
| Applications | Immunofluorescence |
| Storage | 12 months from date of receipt, -20 to -70°C as supplied |
| Background | 180-200 kDa 12 transmembrane member of the Patched family |
| Expression Pattern | High levels in skin and spermatocytes |
Emerging Research Directions
Recent scientific investigations continue to expand our understanding of PTCH2 biology and potential applications for PTCH2 antibodies:
Stem Cell Biology
The 2022 study demonstrating PTCH2's role in regulating mesenchymal stem cells opens new research directions for investigating stem cell maintenance, differentiation, and tissue regeneration . This finding suggests PTCH2 antibodies may become increasingly important tools for stem cell research and regenerative medicine applications.
Tissue-Specific Expression Patterns
Research highlighting PTCH2's restricted expression pattern compared to PTCH1, with particularly high levels in skin and spermatocytes , suggests specialized functions in these tissues. This differential expression pattern indicates potential utility for PTCH2 antibodies in dermatological research and reproductive biology studies.
Development of Therapeutic Applications
As understanding of PTCH2's biological functions continues to expand, particularly regarding its roles in growth control and stem cell regulation, PTCH2 antibodies may find applications beyond basic research into therapeutic development, potentially for conditions involving aberrant cell growth or impaired tissue regeneration.
Product Specs
Target Background
- Combined heterozygous germline mutations in PTCH1 and PTCH2 were identified in a patient with embryonal rhabdomyosarcoma. PMID: 29230040
- Frameshift mutation in the PTCH2 gene can cause nevoid basal cell carcinoma syndrome. PMID: 23479190
- PTCH2 isoforms have distinct roles in Hedgehog signaling. PMID: 14613484
- PTCH2 (2157G-->A), a novel missense mutation, underlies NBCCS, resulting in the loss of PTCH2 inhibitory function in the Shh signaling pathway. PMID: 18285427
- A susceptibility locus on 1p32-1p34 for congenital macrostomia in a Chinese family and identification of a novel PTCH2 mutation are reported. PMID: 19208383
Q&A
What is PTCH2 and what are its primary functions in cellular signaling?
PTCH2 is a 12-transmembrane protein that belongs to the Patched family. Together with Smoothened (SMO), PTCH2 forms a membrane complex that serves as a receptor for Sonic Hedgehog (SHH) . It plays a significant role in the control of cellular growth and may have specific functions in epidermal development . Recent research has demonstrated that PTCH2 is critical in hematopoietic development, with Ptch2 loss driving myeloproliferation and myeloproliferative neoplasm pathogenesis .
Unlike the more ubiquitously expressed PTCH1, PTCH2 has a more restricted expression pattern with particularly high levels found in skin and spermatocytes . The 1182 amino acid mouse Patched 2 contains two terminal cytoplasmic domains (amino acids 1-57 and 1108-1182) and shares 95% amino acid sequence identity with human Patched 2 over the first extracellular loop (amino acids 79-394) .
What are the molecular characteristics of PTCH2 that researchers should be aware of?
PTCH2 is a protein with a molecular weight of approximately 130-145 kDa, as detected in Western blotting applications . The human PTCH2 has several alternative names including UNQ560/PRO1121/PRO57079, PTCH2, Protein patched homolog 2, and PTC2 .
Mouse PTCH2 has a potential isoform that is 1117 amino acids in length and lacks amino acids 207-271 . This variation in isoforms is an important consideration when selecting antibodies and designing experiments, as different epitopes may be present or absent in specific isoforms.
What applications are commercially available PTCH2 antibodies validated for?
The following table summarizes the validated applications for PTCH2 antibodies from different sources:
| Source | Applications | Species Reactivity | Format | Catalog Number |
|---|---|---|---|---|
| Cell Signaling Technology | Western Blotting (1:1000), Immunoprecipitation (1:50) | Human | Rabbit polyclonal | #2464 |
| R&D Systems | Immunohistochemistry | Mouse | Rat monoclonal | MAB8096 |
| Abcam | Western Blotting | Human | Rabbit polyclonal | ab151775 |
When selecting an antibody, researchers should note that reactivity may be limited to specific species, and some antibodies may be suitable only for detecting overexpressed or transfected proteins rather than endogenous levels .
How should researchers optimize PTCH2 detection in immunohistochemistry applications?
For immunohistochemical detection of PTCH2, the protocol should be carefully optimized based on the target tissue and fixation method. For frozen tissue sections, as demonstrated with the R&D Systems antibody, the following methodology has proven effective:
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Fix tissue sections using immersion fixation
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Apply the primary antibody at a concentration of 5 μg/mL
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Incubate overnight at 4°C
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Visualize using an appropriate secondary antibody (e.g., NorthernLights™ 557-conjugated Anti-Rat IgG)
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Counterstain with DAPI for nuclear visualization
This approach has successfully detected PTCH2 in mouse testis sections, which is consistent with the high expression levels reported in spermatocytes . Researchers working with different tissue types should perform titration experiments to determine optimal antibody concentrations.
What are the critical parameters for Western blotting detection of PTCH2?
When performing Western blotting for PTCH2:
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Prepare cell or tissue lysates using standard protocols that preserve membrane proteins
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Load sufficient protein (typically 20-50 μg per lane)
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Use the appropriate dilution of primary antibody (1:1000 for Cell Signaling antibody #2464)
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Include positive controls such as HeLa cell lysate, which has been validated for PTCH2 detection
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Consider longer exposure times if detecting endogenous PTCH2, as expression levels can be low in many cell types
Notably, some PTCH2 antibodies may only detect transfected or overexpressed PTCH2 and not endogenous levels in all cell types .
How does PTCH2 function differ from PTCH1, and what experimental approaches can distinguish between them?
While both PTCH1 and PTCH2 function as receptors in the Hedgehog signaling pathway, they exhibit different expression patterns and potentially distinct functions. PTCH2 is less widely expressed than PTCH1, with high expression primarily in skin and spermatocytes .
To distinguish between PTCH1 and PTCH2 functions experimentally:
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Use specific antibodies that do not cross-react between the two proteins
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Design knockdown or knockout experiments targeting each receptor separately
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Perform rescue experiments with constructs expressing either PTCH1 or PTCH2
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Analyze downstream signaling effects on Gli transcription factors
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Compare phenotypes in tissue-specific conditional knockout models
Research has shown that unlike PTCH1, which is strongly tumor-suppressive, PTCH2 loss specifically affects hematopoietic development, causing myeloproliferation and accelerating JAK2V617F-driven pathogenesis .
What is the role of PTCH2 in hematopoietic development and how can researchers study these effects?
Ptch2 loss drives significant hematopoietic abnormalities, including:
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Leukocytosis with progressive increase in CD11b+ myeloid cells (>60% of peripheral blood at 12 months compared to <20% in controls)
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Massive increase in immature myeloid progenitors (LK cells, 4-fold) and LKS cells (6-fold) in peripheral blood
To study these effects, researchers should consider:
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Flow cytometry analysis of peripheral blood, bone marrow, and spleen populations
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Colony formation assays to assess progenitor function
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Apoptosis assays, particularly for lymphoid populations which show increased rates of spontaneous apoptosis in Ptch2-/- models
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Assessment of specific markers for hematopoietic stem cells and various progenitor populations
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Long-term studies (3, 6, and 12 months) to capture the progressive nature of the phenotype
Why might researchers observe variable or weak PTCH2 detection across different tissue types?
Variable or weak PTCH2 detection can result from several factors:
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Tissue-specific expression patterns: PTCH2 has a more restricted expression profile than PTCH1, with highest levels in skin and spermatocytes
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Antibody sensitivity limitations: Some antibodies may only detect transfected or overexpressed PTCH2
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Protein localization challenges: As a 12-transmembrane protein, PTCH2 may require specialized extraction and preparation methods
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Fixation and processing effects: Improper fixation can mask epitopes or degrade membrane proteins
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Presence of different isoforms: Alternative splicing (such as the mouse isoform lacking aa 207-271) may affect antibody binding
When troubleshooting, researchers should:
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Test multiple antibodies recognizing different epitopes
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Optimize protein extraction protocols for membrane proteins
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Consider using overexpression controls
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Validate results with complementary methods such as qPCR
What are the optimal storage and handling conditions for maintaining PTCH2 antibody activity?
To maintain optimal PTCH2 antibody activity, follow these storage guidelines:
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Store unopened antibodies at -20°C to -70°C for up to 12 months from the date of receipt
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After reconstitution, store at 2-8°C under sterile conditions for up to 1 month
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For longer storage after reconstitution, aliquot and store at -20°C to -70°C for up to 6 months
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Avoid repeated freeze-thaw cycles by preparing appropriate aliquots
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Use manual defrost freezers rather than self-defrosting models to prevent temperature fluctuations
These storage conditions are critical for maintaining antibody performance and preventing degradation that could lead to inconsistent results.
How can PTCH2 antibodies be utilized in co-immunoprecipitation studies to investigate protein-protein interactions?
Co-immunoprecipitation (Co-IP) with PTCH2 antibodies can reveal important protein-protein interactions within the Hedgehog signaling pathway. The Cell Signaling Technology PTCH2 antibody has been validated for immunoprecipitation at a 1:50 dilution .
When designing Co-IP experiments:
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Use gentle lysis buffers that preserve membrane protein complexes
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Pre-clear lysates to reduce non-specific binding
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Include appropriate negative controls (isotype-matched non-specific antibodies)
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Consider crosslinking approaches for transient interactions
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Validate interactions with reciprocal Co-IP experiments
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Confirm results with orthogonal methods such as proximity ligation assays
This approach can be particularly valuable for studying the PTCH2-Smoothened complex formation and its regulation by Sonic Hedgehog binding .
What is known about PTCH2 mutations in disease states, and how can antibody-based approaches contribute to this research?
PTCH2 mutations have been implicated in various disease states, with important roles in hematopoietic disorders. Ptch2 loss drives myeloproliferation and accelerates JAK2V617F-driven myeloproliferative neoplasm pathogenesis .
Antibody-based approaches can contribute to this research through:
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Immunohistochemical profiling of patient samples to assess PTCH2 expression levels
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Flow cytometry analysis of hematopoietic populations to identify alterations in PTCH2 expression
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Phospho-specific antibodies to track downstream signaling changes
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Detection of specific PTCH2 mutations or truncations with epitope-specific antibodies
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Therapeutic targeting studies using blocking antibodies against PTCH2 or its binding partners
Research has shown that Ptch2-/- mice develop a progressive phenotype with leukocytosis, splenomegaly, mobilization of hematopoietic progenitors, and eventual loss of B and T lymphocytes , suggesting important roles in maintenance of normal hematopoiesis.
