GATA29 Antibody
Description
GATA2 Antibodies: Applications and Research Findings
GATA2 is a transcription factor critical for hematopoiesis and immune function. Monoclonal antibodies targeting GATA2 are widely used in research and diagnostics.
Key Applications
Clinical Relevance
GATA2 mutations are linked to hypogammaglobulinemia, NK cell deficiency, and autoimmune diabetes. For example:
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Hypogammaglobulinemia: Patients with GATA2 mutations exhibit low IgG, IgA, and impaired vaccine responses, necessitating IVIG therapy .
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Autoimmune Diabetes: Anti-GAD antibodies (unrelated to GATA2) help diagnose autoimmune diabetes in individuals initially misdiagnosed with type 2 diabetes .
GATA29 in Plant Biology: Gene Regulation and Cold Tolerance
While no antibodies for GATA29 are documented, GATA29 (e.g., SlGATA29 in tomato) interacts with SWIB proteins to suppress WRKY34 expression, influencing cold tolerance.
Mechanistic Insights
Experimental Evidence
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Dual-LUC Assays: Co-transfection of SlGATA29 and SlSWIBa/b reduces WRKY34 promoter activity .
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CRISPR/Cas9 Editing: A 60 bp InDel in the WRKY34 promoter disrupts GATA29/SWIB binding, rendering domesticated tomatoes cold-sensitive .
Table 1: GATA2 Antibody Sensitivity and Immunoglobulin Levels
| Patient | IgG (mg/dL) | IgA (mg/dL) | Vaccine Response |
|---|---|---|---|
| 1 (14 years) | 260 (low) | <7 (undetectable) | Poor response to tetanus/Prevnar vaccines |
| 2 (50 years) | 1122 (on IVIG) | 89 (normal) | Normalized PRP titers post-Hib vaccine |
Source: Adapted from Patient 1 and 2 data in .
Table 2: GATA2 Antibody Detection in Western Blotting
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
Q&A
Here’s a structured collection of FAQs for researchers working with GATA29 Antibody in Arabidopsis thaliana research, integrating technical depth and methodological guidance:
How to validate GATA29 antibody specificity in Arabidopsis mutants?
Method:
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Perform Western blot using knockout (KO) mutants (e.g., T-DNA insertion lines for At3g20750). Compare band presence/absence between wild-type and mutant lysates .
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Combine with immunofluorescence in root or leaf tissues to confirm subcellular localization consistency with known nuclear GATA transcription factors .
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Use peptide competition assays by pre-incubating the antibody with synthetic peptides matching the target epitope (N/C/M terminus) .
What experimental controls are critical for ChIP-seq using GATA29 antibodies?
Controls:
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IgG isotype control to assess non-specific binding.
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Input DNA for normalization.
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Mutant tissue (GATA29 KO) to confirm signal specificity.
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Spike-in DNA (e.g., Arabidopsis-unrelated DNA) for quantitative cross-experiment comparisons .
How to resolve discrepancies in GATA29 protein abundance vs. transcript levels?
Troubleshooting framework:
What engineering strategies improve GATA29 antibody performance in plant tissue homogenates?
Approach:
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Framework humanization: Transplant murine CDRs onto human IgG frameworks to reduce aggregation in acidic plant extracts .
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Glycoengineering: Modify Fc N-glycosylation (e.g., afucosylation) to enhance thermal stability .
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Avidity tuning: Convert to IgM format for low-abundance target detection in complex matrices .
Conflicting subcellular localization results between studies: nuclear vs. cytoplasmic
Resolution workflow:
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Antibody validation: Compare results from N-terminus vs. C-terminus antibodies .
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Conditional analysis: Check stress treatments (e.g., light/dark cycles) affecting GATA29 trafficking.
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Orthogonal validation: Generate GATA29-GFP transgenic lines for live-cell imaging .
Experimental Design Table
| Application | Antibody Region | Recommended Validation | Key Consideration |
|---|---|---|---|
| Western Blot | C-terminus (X-Q9LT45-C) | KO mutant + peptide blocking | Non-specific bands common in plant extracts |
| ChIP-seq | N-terminus (X-Q9LT45-N) | Spike-in DNA + IgG control | Epitope accessibility in crosslinked chromatin |
| ELISA | M-terminus (X-Q9LT45-M) | Standard curve with recombinant protein | Hook effect at high concentrations |
Structural Insights for Advanced Users
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GATA29 antibodies likely use CDR-H3 conformers of length 12–14 residues for target engagement.
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Framework residues at positions 71–78 (VL) are critical for maintaining CDR-L1 geometry in plant-specific epitopes.
