SRM1 Antibody
Product Specs
Composition: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
This antibody targets a transcription activator that regulates abscisic acid (ABA) biosynthesis and signaling. It achieves this by binding to the specific promoter motif 5'- (A/T)AACCAT-3'. Furthermore, it represses ABA-mediated salt stress tolerance (e.g., NaCl and KCl) and influences leaf morphology, promoting vegetative growth.
- Studies have demonstrated opposing roles for MYBS1 and MYBS2 in regulating glucose and abscisic acid signaling during Arabidopsis seed germination and early seedling development. (MYBS2) PMID: 27866313
- At5g08520 functions as a key transcriptional regulator, directly targeting genes involved in abscisic acid biosynthesis and signaling. This highlights its significant role in ABA-mediated salt stress tolerance. PMID: 26243618
Q&A
Basic Research Questions
What validation methods ensure specificity of SRM1 antibodies in mitochondrial studies?
To confirm SRM1 antibody specificity:
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Perform knockout (KO) validation using tissues/cells from SARM1 KO models (e.g., prion-infected mice showing no SARM1 expression in mitochondrial assays) .
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Combine Western blotting and immunohistochemistry (IHC) to verify band size (predicted: ~70 kDa) and subcellular localization (mitochondrial/nuclear) .
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Use pre-adsorption controls with recombinant SRM1 protein to block antigen binding .
How do I optimize SRM1 antibody dilution for immunostaining?
Follow a tiered approach:
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Start with manufacturer-recommended dilutions (e.g., 1:100–1:500 for IHC).
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Test serial dilutions in positive control tissues (e.g., brain thalamus for mitochondrial localization) .
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Validate using secondary antibody-only controls to eliminate background noise .
Table 1: Direct vs. Indirect Immunostaining for SRM1
| Parameter | Direct Method | Indirect Method |
|---|---|---|
| Sensitivity | Low | High |
| Cross-reactivity | Minimal | Requires validation |
| Time | 1–2 hours | 4–6 hours |
| Signal Amplification | None | Yes (via secondary) |
| Adapted from |
Advanced Research Questions
How to resolve contradictory data on SRM1’s role in mitochondrial respiration?
Conflicting reports (e.g., increased mitochondrial respiration in SARM1 KO mice vs. reduced activity in yeast models) require:
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Model-specific validation: Compare antibody performance across species (e.g., murine vs. yeast SRM1 homologs) using cross-reactivity alignment tools (>85% sequence homology recommended) .
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Functional assays: Pair antibody-based detection with mitochondrial respirometry (e.g., Seahorse assays) to correlate protein levels with oxidative activity .
What experimental controls mitigate age-dependent variability in SRM1 localization?
SRM1 nuclear accumulation increases with cellular aging . Address this by:
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Synchronizing cell cycles in cultured models to control for mitotic stage.
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Including age-matched controls in animal studies (e.g., 75 DPI timepoints for prion models) .
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Using dual staining with cell cycle markers (e.g., Ki67) to contextualize localization patterns .
How to validate SRM1 antibody specificity in co-immunoprecipitation (Co-IP) assays?
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Cross-link free vs. cross-linked protocols: Test both to distinguish direct interactions (e.g., SRM1-GSP1 binding) from indirect associations .
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CRISPR-edited lines: Use SRM1 KO cells to confirm loss of co-precipitated targets (e.g., mitochondrial complexes II/IV) .
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Mass spectrometry validation: Compare IP eluates from wild-type and KO models to identify non-specific binders .
Methodological Considerations
How do I address batch-to-batch variability in SRM1 antibodies?
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Request lot-specific validation data from suppliers, focusing on EC50 values in ELISA or neutralization titers .
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Establish an internal reference standard (e.g., a validated lysate) for normalization across experiments .
What assays best correlate SRM1 antibody binding with functional outcomes?
How to troubleshoot non-reproducible SRM1 staining in archival tissues?
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Antigen retrieval: Optimize pH (6.0–9.0) and heating time (20–40 mins) based on fixation method .
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Post-translational modifications: Treat samples with phosphatases/proteases if studying phosphorylation-dependent epitopes .
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Multiplex staining: Combine with mitochondrial markers (e.g., COX IV) to confirm subcellular patterns .
