AE-1435 EzApply 2D Kit
Purpose and Application
- Extraction of Proteins from Mammalian cell or tissue cultures
- 2D electrophoresis sample buffer
Features
- Separate extraction allows higher protein concentration per gel, spot detection is easy.
- Reducing reagent, alkylating reagent, washing buffer and stratified solutions are also incorporated into a kit
- Low running cost, while allowing short-time extraction and easy sample preparation
- Combination use with AE-1430 EzApply (sample preparation kit for SDS-PAGE) makes it applicable to SDS-PAGE as well as 2-D electrophoresis
ATTO Electrophoresis Sample Preparation Reagent Selection Guide
| AE-1430 EzApply | AE-1435 EzApply 2D kit | WSE-7010 EzLabel FluoroNeo | WSE-7011 EzApply Native | WSE-7040 EzApply DNA |
|
|---|---|---|---|---|---|
| Application | SDS-PAGE sample preparation (2×) | Sample preparation for two-dimensional electrophoresis | SDS-PAGE sample preparation | Native-PAGE sample preparation (10×) | DNA electrophoresis loading dye (6×) |
| Main components | Tris-HCl buffer, SDS, glycerol, DTT, BPB | ① Wash buffer: Tris, NaCl ② Solution 1: Tris, ampholytes, DTT ③ Solution 2: urea, thiourea, detergents, ampholytes, DTT ④ Solution 1-2: powder ⑤ Solution 2-2: powder ⑥ Solution 1 concentrated solution: BPB ⑦ Solution 2 concentrated solution: urea | ① Sample buffer (5×): detergents, buffer components ② Labeling reagent: fluorescent dye ③ Reducing agent (DTT): DTT ④ BPB (bromophenol blue) ⑤ Molecular weight marker: proteins/glycerol ⑥ RIPA lysis buffer: detergents, buffering agents | Tris, glycerol, bromophenol blue, xylene cyanol | 15% Ficoll, 0.05% bromophenol blue, 0.05% red dye |
| Volume / Contents | 5 mL × 5 | ① Wash buffer: 30 mL × 2 ② Solution 1: 20 mL ③ Solution 2: 10 mL ④ Solution 1-2: powder ⑤ Solution 2-2: powder ⑥ Solution 1 conc.: 1 mL ⑦ Solution 2 conc.: 1 mL ⑧ DTT for Solution 1: powder ⑨ DTT for Solution 2: powder | ① Sample buffer (5×): 12 mL ② Labeling reagent: 10 mg ③ Reducing agent (DTT): 300 mg ④ MW marker: 600 µL (220, 169, 97, 66, 45, 30, 20.1, 14.4 kDa) ⑤ RIPA lysis buffer: 10 mL | 40 mL | 10 mL |
| Preparation | Prepare 2× stock solution: add 5 mL EzApply to the DTT bottle | Prepare by adding DTT to Solution 1 or Solution 2 | Dissolve labeling reagent in MeOH or acetonitrile; dissolve DTT in DW | Not required | Not required |
| Usage | Mix prepared EzApply with protein sample at 1:1 ratio; boil for 5 min | Preparation method varies depending on sample; refer to the instruction manual | Add sample buffer and labeling reagent to sample, heat at 95 °C for 3 min; then add DTT solution and heat again at 95 °C for 3 min | Add at 1/10 volume to protein extract | Add 1 µL of this reagent to 5 µL of DNA solution |
| Capacity | 1,250 samples (when using 20 µL per 20 µL sample) | ~20 extractions from 100 mg mammalian tissue or cells | 2,000 samples (when using 5 µL per 20 µL sample) | As required | As required |
| Storage | −20 °C, 6 months | −20 °C, 6 months | −20 °C, 1 year | Refrigerated, 1 year | Refrigerated, 1 year |
Instruction Manual
Specifications
| AE-1435 EzApply 2D Kit | |
|---|---|
| Major components | Wash buffer: Tris, NaCl Solution 1: Tris, Ampholine, DTT Solution 2: Urea, Thiourea, Surfactant Alkylated reagent: Ampholine, DTT Stratified solutions: Acrylamide, BPB, Urea |
| Package | Solution/30 mL bottle x 2 (Wash buffer) Solution/20 mL bottle (Solution 1) Solution/10 mL bottle (Solution 2) Solution/1 mL tube (Alkylated reagent) Solution/500 μL tube (Stratified solution) Powder/tube (DTT) |
| Preparation | Appropriated amount of solution is applied to tissues or cells, which were lysed and extracted |
| Applicable amount | Extracted from 100 mg of mammalian tissue 20 runs |
| Storage | 6 months at -20℃ (before dissolved in DTT) 1 week after dissolved in DTT |
Ordering Information
| Code No. | Description | Unit |
|---|---|---|
| 2332335 | AE-1435 EzApply 2D Kit | 1 pk |
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