CRK15 Antibody
Description
CRK Family Antibodies (CT10 Regulator of Kinase)
The CRK protein family (cellular homolog of the v-Crk oncogene) contains two main isoforms (Crk-I and Crk-II) and the related CRKL protein. Antibodies targeting these proteins show significant research applications:
Potential Typographical Confusion
The term "CRK15" may represent a conflation of:
A) CRK + Cytokeratin 15 (KRT15)
| Parameter | CRK Antibodies | Cytokeratin 15 Antibodies |
|---|---|---|
| Target Protein | Adaptor signaling proteins | Epithelial structural protein (KRT15) |
| Key Applications | Cancer signaling studies | Stem cell/tissue regeneration research |
| Commercial Example | MA5-15891 (Thermo Fisher) | CAB4854 (Assay Genie) |
| Validation Data | Apoptosis assays, motility studies | IHC, flow cytometry |
B) CRKL Isoforms
CRKL (CRK-like protein) shares structural homology with CRK but contains distinct SH2/SH3 domains. Antibodies like ABIN969062 show cross-reactivity with CRKL across human, monkey, mouse, and rat samples ( ).
Technical Validation Data
From CRK-related antibody studies:
Functional Epitopes
| Antibody Clone | Epitope Region | Biological Impact |
|---|---|---|
| 3G11C1 (MA5-15891) | Recombinant human CRK | Inhibits Rac-dependent cell motility |
| shCRKL#3 (H1755) | CRKL 3′UTR | Induces PARP/Caspase-3 cleavage |
Product Specs
**Constituents:** 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
Q&A
Basic Research Questions
How to validate CRK15 antibody specificity in immunohistochemistry (IHC)?
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Perform knockout validation using CRISPR-edited cell lines (e.g., KRT15 knockout A431 cells) to confirm loss of signal compared to wild-type controls .
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Use multiple antibody clones (e.g., [LHK15] vs. [EPR1614Y]) to cross-verify staining patterns in tissues like normal breast or cervical carcinoma .
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Include isotype controls and pre-absorption with recombinant CRK15 protein to rule out nonspecific binding .
What are common applications of CRK15 antibodies in epithelial cell research?
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Cell lineage tracing: Detect stem cell populations in stratified epithelia (e.g., hair follicles, mammary glands) using paraffin-embedded tissues .
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Tumor subtyping: Differentiate basal-like carcinomas (CRK15+) from luminal subtypes in breast cancer studies .
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Mechanical stress response assays: Monitor cytoskeletal reorganization in keratinocytes under shear stress .
How to optimize CRK15 detection in Western blotting?
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Use heat-mediated antigen retrieval (citrate buffer, pH 6.0) and reduce sample heating to prevent epitope degradation .
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Load 20 µg/lane of lysate from tissues like mouse skin or thymus for clear detection of the ~50 kDa band .
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Validate with recombinant full-length CRK15 protein as a positive control .
Advanced Research Questions
How to resolve contradictory CRK15 staining results across studies?
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Statistical reconciliation: Apply Cohen’s kappa analysis to inter-observer variability in IHC scoring .
What mechanisms link CRK15 to oncogenic pathways like RAS/EGFR?
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CRK15 interacts with RAP1 GTPase to promote EGF-independent proliferation in NSCLC (via CRKL amplification) .
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In Drosophila models, CRK family proteins regulate Yki/YAP oncogenic activity through JNK-mediated F-actin accumulation .
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Co-expression networks: Use multiplex IHC to map CRK15 with EGFR or p38 MAPK in tumor microarrays .
How to design CRK15 functional studies in 3D organoid models?
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Step 1: Generate air-liquid interface cultures of primary keratinocytes .
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Step 2: Transduce with CRK15 shRNA (e.g., doxycycline-inducible systems) .
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Step 3: Quantify spheroid invasion using collagen-embedded assays and live-cell imaging .
