CBL10 Antibody
Description
CBL10 Protein Overview
CBL10 is a calcium-binding protein in the calcineurin B-like (CBL) family that decodes calcium signals during abiotic stress responses. It interacts with CBL-interacting protein kinases (CIPKs) to regulate ion transport and cellular signaling .
A. Salt Tolerance Pathway
CBL10 partners with CIPK24 (SOS2) to mediate Na⁺ sequestration into vacuoles under salt stress. This pathway is distinct from the plasma membrane-localized SOS3-CIPK24 system :
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Mutant Phenotype: cbl10 mutants exhibit hypersensitivity to salt stress, with reduced Na⁺ accumulation in shoots .
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Electrophysiology: CBL10-CIPK24 inhibits AKT1-mediated K⁺ uptake in root cells, redirecting resources to Na⁺ detoxification .
B. Interaction with AKT1
CBL10 directly binds the potassium channel AKT1, modulating its activity:
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Functional Impact: Overexpression of CBL10 reduces AKT1-mediated K⁺ currents, altering ion balance under low-K⁺ conditions .
C. GTPase Regulation
CBL10 inhibits TOC34 (a chloroplast protein import GTPase) in a Ca²⁺-dependent manner, suggesting a role in chloroplast signaling :
Comparative Roles of CBL Proteins
A. Agricultural Applications
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Salt Tolerance Engineering: Overexpression of CBL10 could enhance crop resilience to saline soils .
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Ion Homeostasis: Targeting CBL10-CIPK24 interactions may optimize nutrient uptake under stress .
B. Unresolved Questions
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Downstream Targets: The Na⁺ transporter activated by CBL10-CIPK24 remains unidentified .
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Cross-Species Relevance: Whether CBL10 homologs in crops (e.g., rice, wheat) share similar functions requires study.
Antibody Development Context
While no commercial "CBL10 Antibody" is listed in the reviewed sources, antibodies against related CBL proteins (e.g., CBLB in humans) use epitopes in conserved regions . For example:
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CBL Antibody (A-9): Targets the C-terminus of human/mouse CBL (aa 892–906) .
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Validation Methods: Western blot, IP, and immunofluorescence .
Future Directions
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Antibody Generation: Designing antibodies against Arabidopsis CBL10’s unique C-terminal domain (aa 892–906 homolog) could aid in subcellular localization studies.
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Therapeutic Potential: In humans, CBLB inhibitors (e.g., benzodiazepines) enhance antitumor immunity by activating T-cells , suggesting parallels in plant immune modulation via CBL10.
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
The CBL10 antibody targets a calcium sensor protein. CBL proteins interact with CIPK serine-threonine protein kinases. Calcium-dependent binding of a CBL protein to the regulatory NAF domain of a CIPK protein activates the kinase. CBL10 mediates salt tolerance, but only in its phosphorylated state. It also negatively regulates AKT1 activity through a protein kinase-independent mechanism, competing with CIPK23 for AKT1 binding.
CBL10 Function and Regulation: Research indicates CBL10 plays a multifaceted role in plant development and stress response. Key findings include:
- Reproductive Development and Salt Stress: CBL10 is crucial for reproductive development under saline conditions and functions differently during vegetative and reproductive growth. (PMID: 26979332)
- Drought Tolerance and Brassinosteroid Sensitivity: CBL10 negatively impacts drought tolerance, independent of abscisic acid signaling. Calcium signaling via CBL10 differentially affects abiotic stress responses, partially through modulating brassinosteroid sensitivity. (PMID: 26795150)
- Salt Tolerance and Transcriptional Regulation: Studies suggest a negative regulatory role of ANT on SCABP8 under salt stress, as indicated by the suppression of the salt-tolerant phenotype in ant knockout mutants by SCABP8 mutation. (PMID: 26054800)
- AKT1 Regulation and Ion Homeostasis: CBL10 directly regulates AKT1 activity, influencing ion homeostasis through a CBL-interacting protein kinase-independent mechanism. (PMID: 23331977)
- SCABP8 and SOS3 Redundancy: SCABP8 and SOS3 exhibit partial functional redundancy, each contributing unique roles to the plant's salt stress response. (PMID: 17449811)
- Novel Salt Tolerance Pathway: CBL10 and CIPK24 (SOS2) form a novel salt-tolerance pathway regulating sodium sequestration/compartmentalization. (PMID: 17825054)
- SCaBP8 Phosphorylation and SOS1 Activation: SCaBP8 phosphorylation is essential for SOS1 activation and salt tolerance regulation in Arabidopsis. (PMID: 19448033)
Q&A
Clarification: The provided search results focus on CD10/CALLA antibodies (e.g., SHB-10, eBioCB-CALLA) rather than "CBL10 Antibody." Assuming this discrepancy arises from a nomenclature variation or typographical error, the FAQs below address CD10/CALLA antibodies, which are well-documented in the literature. If "CBL10" refers to a distinct target, additional data would be required.
How do I validate a CD10/CALLA antibody for flow cytometry in acute lymphoblastic leukemia (ALL) studies?
Methodological Answer:
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Epitope Specificity: Confirm antibody binding to the 100–105 kDa CD10 glycoprotein via immunoprecipitation or Western blot under reducing/non-reducing conditions .
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Blocking Assays: Use competitive binding assays (e.g., pre-incubation with recombinant CD10) to verify specificity. SHB-10 blocked anti-CD10 binding to Daudi cells and ALL lymphocytes in flow cytometry .
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Cell Line Controls: Test reactivity against CD10-positive (e.g., ALL cell lines) and CD10-negative cell lines (e.g., non-lymphoid tumors) .
Table 1: Key Validation Parameters for CD10 Antibodies
| Parameter | Example from Literature | Source |
|---|---|---|
| Molecular Weight | 100–105 kDa (reduced conditions) | |
| Blocking Efficiency | SHB-10 blocks >80% of anti-CD10 binding | |
| Cell Reactivity | Daudi cells, ALL lymphocytes |
What are the primary technical applications of CD10 antibodies in biomedical research?
Methodological Answer:
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Flow Cytometry (FCM): Use CD10 antibodies (e.g., eBioCB-CALLA) at ≤0.5 µg/test for immunophenotyping ALL samples .
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Immunohistochemistry (IHC): Target CD10 in renal tubules, gut epithelia, or germinal center B cells with antibodies validated for paraffin-embedded tissues .
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Functional Assays: Study CD10’s metallopeptidase activity by measuring cleavage of substrates like bradykinin or neurotensin in enzymatic assays .
Note: Antibodies against linear epitopes (e.g., anti-peptide) are better for Western blot, while conformation-specific antibodies are ideal for FCM .
How to resolve discrepancies in CD10 expression profiles across neuroectodermal tumor cell lines?
Methodological Answer:
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Surface vs. Intracellular Staining: Use flow cytometry with/without permeabilization to distinguish membrane-bound vs. cytoplasmic CD10 .
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Orthogonal Validation: Combine flow cytometry with mass spectrometry or RNA sequencing to correlate protein and mRNA levels .
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Context-Specific Optimization: Adjust fixation protocols (e.g., methanol-free for epitope preservation) to mitigate artifacts .
Example: SHB-10 showed slight differences in neuroectodermal tumor cell surface expression compared to conventional anti-CD10, suggesting epitope accessibility varies by cell type .
How to design controls for CD10 antibody specificity in multiplex assays?
Methodological Answer:
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Isotype Controls: Use same-host IgG with identical fluorophore conjugation .
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Knockout Validation: CRISPR-Cas9 CD10-knockout cell lines to confirm signal loss .
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Cross-Reactivity Screening: Test antibody against off-target proteins (e.g., CD13, CD26) using protein microarrays .
Table 2: Common Pitfalls in CD10 Antibody Applications
How to interpret conflicting data on CD10’s role in neutrophil chemotaxis?
Methodological Answer:
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Functional Blocking: Use neutralizing CD10 antibodies (e.g., eBioCB-CALLA) to inhibit metallopeptidase activity and measure chemokine gradients (e.g., IL-8) .
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Dynamic Assays: Perform live-cell imaging with fluorescent substrates (e.g., FITC-bradykinin) to quantify cleavage kinetics .
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Meta-Analysis: Compare datasets from antibody-dependent studies (e.g., Human Protein Atlas) with genetic knockout models .
Key Finding: CD10 regulates stromal-dependent B lymphopoiesis but may exhibit context-dependent roles in inflammation due to substrate competition .
