LCR22 Antibody
Description
LCR22 (Low Copy Repeats on Chromosome 22)
LCR22 refers to low copy repeats on chromosome 22q11.2, which mediate genomic rearrangements linked to disorders like 22q11.2 deletion syndrome (DiGeorge syndrome). These regions are not direct targets for antibodies but are instead structural genomic elements.
CD22 (Cluster of Differentiation 22)
CD22 is a B-cell surface glycoprotein encoded by a gene on chromosome 19q13.12. Antibodies targeting CD22 are used in immunotherapy for B-cell malignancies.
CD22-Targeted Antibodies in Clinical Development
While no antibodies directly target LCR22, multiple anti-CD22 antibodies and CAR T-cell therapies are under investigation. Key examples include:
Anti-CD22 Monoclonal Antibodies
CD22 CAR T-Cell Therapies
Binding Epitopes
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m971: Targets domains 5–7 of CD22, distinct from earlier antibodies (e.g., HA22 targeting domain 3) .
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HA22: Derived from murine RFB4; binds domain 3 with partial competition .
Functional Properties
| Property | m971 | HA22 |
|---|---|---|
| Affinity (EC50) | 2 nM (Fab), <1 nM (IgG1) | 20 nM (Fab) |
| Internalization | Rapid (Raji cells) | Moderate |
| Clinical Safety | No grade >3 CRS/ICANS | Limited by immunogenicity |
Challenges and Future Directions
Product Specs
Constituents: 50% Glycerol, 0.01M Phosphate Buffered Saline (PBS), pH 7.4
Target Background
KEGG: ath:AT4G29280
STRING: 3702.AT4G29280.1
Q&A
What is CCRL2/LCCR and how is its antibody used in research?
CCRL2 (Chemokine Receptor-Like 2), also known as LCCR (Lipopolysaccharide-inducible CC chemokine receptor), is a 7-transmembrane domain receptor expressed on macrophages, glial cells, and mast cells at inflammatory sites. It binds chemerin and CCL2, 5, 7, and 8, mediating immune responses . The anti-CCRL2 antibody (e.g., Rat Anti-Mouse CCRL2/LCCR APC-conjugated Monoclonal Antibody, Clone #498321) is primarily used for:
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Flow cytometry: Detecting CCRL2 expression on immune cells (e.g., J774A.1 macrophage cell line) .
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Immunofluorescence: Localizing CCRL2 in tissue sections.
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Validation: Confirming receptor presence in experimental models.
What cell types express CCRL2 and how to validate antibody specificity?
CCRL2 is expressed on activated immune cells, including macrophages, glial cells, and mast cells . To validate antibody specificity:
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Use positive controls: Test on known CCRL2+ cell lines (e.g., J774A.1 macrophages) .
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Include isotype-matched controls: Compare binding to non-specific IgG (e.g., Catalog #IC005A) .
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Adopt ELISA-based methods: Immobilize cell membrane proteins (1 mg/mL concentration) and assess antibody binding via OD readings, as demonstrated for anti-CD22 antibodies .
What experimental applications are suitable for CCRL2 antibodies?
How to optimize flow cytometry protocols for detecting CCRL2 expression?
Key Parameters to Optimize:
Example Workflow:
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Stain cells: Use Rat Anti-Mouse CCRL2 APC (FAB5519A) at 1:100 dilution.
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Compare to isotype: Use IC005A (non-specific rat IgG) to subtract background .
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Gating strategy: Gate on viable, single-cell populations before analyzing CCRL2+ subsets.
How to address cross-reactivity when using anti-CCRL2 antibodies across species?
Strategies:
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Sequence alignment: Mouse CCRL2 shares 49% identity with human and 70% with rat . Avoid cross-species use unless confirmed.
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Blocking peptides: Pre-incubate antibodies with CCRL2-derived peptides to confirm epitope specificity.
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Knockout controls: Validate antibody reactivity in CCRL2-deficient cell lines or mice.
How to interpret flow cytometry data showing variable CCRL2 expression?
Troubleshooting Table:
What methods validate antibody specificity beyond manufacturer claims?
Advanced Validation Approaches:
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Peptide competition: Incubate antibodies with CCRL2-derived peptides to block binding.
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CRISPR knockout models: Confirm loss of signal in CCRL2-deficient cells.
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Orthogonal assays: Compare flow cytometry results with qPCR for CCRL2 mRNA or Western blot (if epitope is denatured).
How to design an experiment to study CCRL2’s role in inflammation using the antibody?
Experimental Framework:
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Model Selection: Use LPS-induced inflammation in mice or in vitro macrophage differentiation.
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Cell Tracking:
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Data Analysis: Correlate CCRL2 expression with cytokine levels (e.g., TNF-α, IL-6) via multiplex assays.
Storage and Handling
Addressing Data Contradictions
Scenario: Anti-CCRL2 antibodies show inconsistent binding in ELISA vs. flow cytometry.
Analysis:
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Epitope accessibility: ELISA detects denatured epitopes, while flow cytometry targets native conformations .
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Cell state: CCRL2 expression may require activation (e.g., LPS stimulation) .
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Buffer compatibility: Optimize blocking agents (e.g., BSA vs. FBS) for each assay.
Single-Cell Analysis
Approach:
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Multi-omic profiling: Combine CCRL2 detection with RNA sequencing to link receptor expression to inflammatory pathways.
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Spatial mapping: Use imaging mass cytometry to localize CCRL2+ cells in inflamed tissues.
