View Featured Offers >>
7723
Red Loading Buffer Pack
Buffers & Dyes
Buffer Kit

Red Loading Buffer Pack #7723

Citations (0)
To Purchase # 7723
Cat. # Size Qty. Price
7723S
1 Pack

Product Includes Quantity
3X Red Loading Buffer 8 ml
30X Reducing Agent (1.25 M DTT) 1 ml

Product Usage Information

1. Prepare fresh 3X Reducing SDS Loading Buffer by adding 1/10 volume 30X DTT Reducing Agent to 1 volume of 3X SDS Loading Buffer.

2. Dilute 3X SDS Loading Buffer to a 1X solution using ddH2O. This product supplies enough 3X material to make 24ml of 1X solution.     

3. Aspirate media from cultures; wash cells with 1X PBS; aspirate.

4. Lyse cells by adding 1X SDS Loading Buffer (100 µl per well of 6-well plate or 500 µl per plate of 10 cm2 plate). Immediately scrape the cells off the plate and transfer the extract to a microcentrifuge tube. Keep on ice.

5. Sonicate for 10-15 seconds to shear DNA and reduce sample viscosity. 

6. Heat a 20 µl sample to 95-100ºC for 5 minutes; cool on ice.

7. Microcentrifuge for 2-5 minutes.

8. Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).

Solutions and Reagents

Quantity 8 ml 3X Red Loading Buffer and 1 ml 30X Reducing Agent3X Red Loading Buffer Pack: 187.5 mM Tris-HCl (pH 6.8 at 25ºC), 6% (w/v) SDS, 30% glycerol and 0.03% (w/v) phenol red. (Store at room temperature.)       30X Reducing Agent: 1.25 M dithiothreitol (DTT) (Store at -20ºC.)

Storage

Store at Room Temperature and -20°C.
3X Red Loading Buffer: 187.5 mM Tris-HCl (pH 6.8 at 25°C), 6% (w/v) SDS, 30% glycerol and 0.03% (w/v) phenol red. (Store at room temperature.)
30X Reducing Agent: 1.25 M dithiothreitol (DTT) (Store at -20°C.)

Product Description

The Red Loading Buffer Pack is used to lyse cells for western blotting analysis.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

For Research Use Only. Not for Use in Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit our Trademark Information page.