2A3
Based on 1 Customer Validation
2A3 (2-aminopyridine-3-carboxylic acid imidazolide) is a covalent probe reagent for Selective 2'-Hydroxyl Acylation analyzed by Primer Extension (SHAPE) that targets the RNA ribose 2'-OH group. 2A3 efficiently permeates the biological membranes of Gram-negative bacteria, Gram-positive bacteria, and mammalian cells. 2A3 exhibits no base bias and specifically labels conformationally flexible, unpaired regions within RNA. 2A3 forms covalent adducts by acylating the 2'-OH groups of flexible RNA residues. When combined with SHAPE-MaP (mutational profiling) sequencing technology, these modification sites are converted into detectable mutational signals, thereby accurately reflecting local RNA backbone flexibility and base-pairing status. 2A3 is primarily utilized in molecular biology and transcriptomics research, including the resolution of RNA structuromes in living cells, the study of RNA folding regulatory mechanisms, and the investigation of non-coding RNA functions.
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- Purity : 99.92%
- CAS No.: 2765091-45-8
- 화학식: C9H8N4O
- 분자량:188.19
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보관:
4°C, protect from light, stored under nitrogen
* In solvent : -80°C, 6 months; -20°C, 1 month (protect from light, stored under nitrogen)
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Tel: 609-228-6898 E-mail: [email protected] [email protected]
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Biological Activity
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Chemical Information
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용액&용해도
- Protocol
- 순도&문서
- References
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Help & FAQs
All DNA/RNA Synthesis Isoforms
More
Biological Activity
제품 설명
In Vitro
In in vitro SHAPE structural probing assays using deproteinized E. coli rRNA, 2A3 (100 mM; 37°C; 15 min) demonstrates significantly higher reactivity toward RNA 2'-hydroxyl groups compared to the classic SHAPE reagent NAI; the median mutation frequency was approximately 2.4 times that of NAI, and the signal-to-noise ratio for distinguishing between single-stranded and double-stranded RNA was superior[1].
In RNA structural probing experiments involving live E. coli, B. subtilis, and HEK293 cells, 2A3 (100 mM; 37°C; 15-20 min) exhibits membrane permeability far superior to that of NAI; the AUC for distinguishing single-stranded from double-stranded RNA in solvent-exposed regions within live cells improved by 17%-20% compared to NAI[1].
In cell integrity assays using HEK293 cells, E. coli, and B. subtilis, 2A3 (100 mM; 37°C; 30 min) does not induce morphological abnormalities, cell lysis, or a decrease in bacterial culture optical density, confirming that its high probing efficiency did not result from non-specific effects associated with cell rupture[1].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only. Further protocols information, click here.
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Cell Line:HEK293, Escherichia coli K-12 DH10B, Bacillus subtilis 168
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Concentration:100 mM
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Incubation Time:30 min at 37°C
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Result:Showed no detectable morphological changes or cell lysis against HEK293 cells.
For both E. coli and B. subtilis, the optical density of the bacterial cultures did not decrease over the incubation period.
Did not disrupt cell integrity, and high probing efficiency was not attributed to non-specific cell lysis.
Chemical Information
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CAS No. 2765091-45-8
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Appearance Solid
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분자량 188.19
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화학식 C9H8N4O
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Color White to off-white
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SMILES
O=C(C1=CC=CN=C1N)N2C=NC=C2
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Synonyms
2-aminopyridine-3-carboxylic acid imidazolide
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선적
Room temperature in continental US; may vary elsewhere.
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보관
4°C, protect from light, stored under nitrogen
* In solvent : -80°C, 6 months; -20°C, 1 month (protect from light, stored under nitrogen)
용액&용해도
In Vitro:
DMSO : 20 mg/mL (106.28 mM; ultrasonic and warming and heat to 60°C; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light, stored under nitrogen). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light, stored under nitrogen). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
In Vivo:
Select the appropriate dissolution method based on your experimental animal and administration route.
- For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
- To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for In Vivo experiments, it is recommended to prepare freshly and use it on the same day.
- The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.
Add each solvent one by one: 10% DMSO 90% (20% SBE-β-CD in Saline)
Solubility: ≥ 2 mg/mL (10.63 mM); Clear solution
This protocol yields a clear solution of ≥ 2 mg/mL (saturation unknown).
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (20.0 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C, storage for one week): 2 g SBE-β-CD powder is dissolved in 10 mL Saline, completely dissolve until clear.
In Vivo Dissolution Calculator
Please enter the basic information of animal experiments:
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Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Please enter your animal formula composition:
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%DMSO +
Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
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%+
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+%Tween-80 + +
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%Saline +
The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).
Working solution concentration: 0.22 mg/mL
Method for preparing stock solution: mg drug dissolved in μL DMSO. Stock solution concentration: mg/mL. * In solvent : -80°C, 6 months; -20°C, 1 month (protect from light, stored under nitrogen)
1. Take μL DMSO stock solution;
2. Add μL .
μL , mix evenly;
3. Then add μL Tween 80, mix evenly;
4. Then add μL
Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.
Protocol
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Research Protocol for Omics Data Analysis Pipelines
Omics data analysis pipelines convert raw high-throughput measurements from genomics, transcriptomics, epigenomics, proteomics, metabolomics, or single-cell assays into quality-controlled, statistically tested, biologically interpretable results. A reproducible omics pipeline requires predefined experimental metadata, raw-data quality control, modality-specific preprocessing, normalization, statistical modeling, multiple-testing correction, biological annotation, and independent validation. RNA-seq pipelines commonly include read QC, alignment or pseudoalignment, quantification, normalization, and differential-expression testing, while single-cell pipelines additionally require cell-level QC, normalization, dimensionality reduction, clustering, cell annotation, and sample-aware differential testing. Multi-omics integration can connect molecular layers such as transcriptome, proteome, metabolome, and epigenome, but unresolved problems include batch effects, missing values, unequal featu
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Gram Staining of Tissue Sections
Gram staining of tissue sections is a histochemical technique used to differentiate Gram-positive and Gram-negative bacteria within histological specimens based on differences in bacterial cell wall structure and dye retention, adapted from classical bacteriological Gram staining into tissue-compatible “histological Gram stain” variants. In tissue applications, modifications of the Brown-Hopps and Brown-Brenn methods are commonly used to improve differentiation of microorganisms embedded within host connective tissue and to reduce overstaining or loss of Gram-negative signal, which are known limitations of earlier approaches. The principle relies on crystal violet-iodine complex retention in Gram-positive organisms and subsequent decolorization and counterstaining steps that allow contrast visualization of Gram-negative organisms against tissue background.
순도&문서
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Data Sheet (278 KB)
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SDS (254 KB)
- English - EN (254 KB)
- Français - FR (254 KB)
- Deutsch - DE (254 KB)
- Norwegian - NO (254 KB)
- Español - ES (254 KB)
- Swedish - SV (254 KB)
- Italian - IT (254 KB)
- Korean - KR (254 KB)
- Portuguese - PT (254 KB)
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Handling Instructions (2659 KB)
References
Complete Stock Solution Preparation Table
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light, stored under nitrogen). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
| Optional Solvent | Concentration Solvent Mass | 1 mg | 5 mg | 10 mg | 25 mg |
|---|---|---|---|---|---|
| DMSO | 1 mM | 5.3138 mL | 26.5689 mL | 53.1378 mL | 132.8445 mL |
| 5 mM | 1.0628 mL | 5.3138 mL | 10.6276 mL | 26.5689 mL | |
| 10 mM | 0.5314 mL | 2.6569 mL | 5.3138 mL | 13.2844 mL | |
| 15 mM | 0.3543 mL | 1.7713 mL | 3.5425 mL | 8.8563 mL | |
| 20 mM | 0.2657 mL | 1.3284 mL | 2.6569 mL | 6.6422 mL | |
| 25 mM | 0.2126 mL | 1.0628 mL | 2.1255 mL | 5.3138 mL | |
| 30 mM | 0.1771 mL | 0.8856 mL | 1.7713 mL | 4.4281 mL | |
| 40 mM | 0.1328 mL | 0.6642 mL | 1.3284 mL | 3.3211 mL | |
| 50 mM | 0.1063 mL | 0.5314 mL | 1.0628 mL | 2.6569 mL | |
| 60 mM | 0.0886 mL | 0.4428 mL | 0.8856 mL | 2.2141 mL | |
| 80 mM | 0.0664 mL | 0.3321 mL | 0.6642 mL | 1.6606 mL | |
| 100 mM | 0.0531 mL | 0.2657 mL | 0.5314 mL | 1.3284 mL |
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