- 询价
- 2900
- 2025年12月18日
- Protein Extraction
- 0
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- 详细信息
- 询价记录
- 文献和实验
- 技术资料
- 应用范围：
Protein Extraction
- 宿主：
0
- 抗原来源：
0
- 库存：
大量
- 是否单克隆：
0
- 规格：
100 assays
For Research Use Only; Not for use in diagnostic procedures.
- Cytoplasmic Lysis Buffer, 10x: - (Part No. 90497) One vial containing 10mL of a concentrated lysis buffer. Dilute to a 1x in deionized water.
- Nuclear Extraction Buffer: - (Part No. 90498) One vial containing 50mL of a 1x nuclear extraction buffer.
- PBS Packets: - (Part No. 60093) Two pouches containing enough dry reagent to prepare 1 liter of a 1x PBS solution per pouch. Dilute packet in deionized water.
- DTT, 1M: - (Part No. 90499) One vial containing 100 μL of 1M Dithiothreitol. Prior to use, dilute to a final concentration of 0.5mM (1:2000) in 1x Cytoplasmic Lysis Buffer and 1x Nuclear Extraction Buffer.
- Protease Inhibitor Cocktail: - (Part No. 90492) One vial containing 100μL of protease inhibitors in DMSO for use with mammalian cell and tissue extract buffers. A mixture of protease inhibitors with broad specificity for the inhibition of serine, cysteine and aspartic acid proteases and aminopeptidases. Contains 4-(2-aminoethyl)benzenesulfonyl fluoride (AEBSF), pepstatin A, E-64, bestatin, leupeptin, and aprotinin. Contains no metal chelators (e.g. EDTA, EGTA). Prior to use, dilute 1/1000 in 1x Cytoplasmic Lysis Buffer and 1x Nuclear Extraction Buffer.
- Detergent, 10%: - (Part No. 90500) One vial containing 3mL of 10% IGEPAL CA-630 (detergent). Cytoplasmic Lysis Buffer and Nuclear Extraction Buffer already contain detergent, however, under certain conditions, more detergent may be required - refer to Extraction Procedure.
·Cytoplasmic Lysis Buffer (10x), Nuclear Extraction Buffer (1x), 10% Detergent, and the PBS Packets can be stored at 2-8ºC.
·DTT, 1M and Protease Inhibitor Cocktail must be stored at -20ºC. Avoid repeated freeze-thaw cycles.
2. Cell Detachment Buffer (Trypsin) or Cell Scrapers
3. Syringes, 1mL - 27 Gauge Needle
4. 1.5 mL Microcentrifuge Tubes
5. 50 mL Conical Tubes
6. Deionized Water
7. Phosphatase Inhibitor (if needed)
8. Microcentrifuge, 4°C
9. Table-top Centrifuge (capable of 16,000 x g), 4°C
10. Rotator/Orbital Shaker, 4°C
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详细描述见链接：http://www.millipore.com/catalogue/item/2900
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文献和实验PMSF (1 mM) Protease Inhibitor Cocktail, Sigma (Cat. no. P-2714) Quant-iT™ Protein Quantitation Kit (Cat. no. Q33210) Or Cell Extraction Buffer Formulation 100 mM Tris, pH 7.4 2 mM Na3 VO4 100 mM NaCl
8.0. Resuspend the final washed nuclear pellet in ice cold distilled water to a final volume of 100 ml and allow to swell overnight at 4° C. Gently stir the mixture on the following day. This solution is the pure chromatin to be used
Preparation of Hela Cell Nuclear and Cytosolic S100 Extracts for In Vitro Splicing
in relatively efficient and accurate splicing, and is now in wide use, with slight variations from laboratory to laboratory. Variations in extract preparation include primarily the use of slightly different buffers and salts for nuclear extraction or dialysis.
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