MES SDS Buffer Kit – For Protein Sample Preparation & Electrophoresis on Bis-Tris Gel

DESCRIPTION

MES SDS Buffer Kit is a complete buffer kit to resolve small- to medium-sized proteins under denaturing conditions on Bis-Tris gels.

The MES SDS Buffer Kit contains the following:

Component	Cat. No. KBMES86-20X-250ML	Cat. No. KBMES86-20X-500ML
MES SDS Running Buffer [20X]	1 x 250 mL	1x 500 mL
10X DTT (500 mM Reducing Agent). Provided as solid powder	1x 50µL (reconstitute with 50µL Molecular Grade Water	1x 100µL (reconstitute with 100µL Molecular Grade Water
Protein Antioxidant [200X]	1x 7.5 mL	1x 15 mL
LDS Sample Loading Buffer [4X]	1x 5 mL	1x 10 mL

The Kit is formulated for protein sample preparation and electrophoretic separation on hand-made or commercially available precast Bis-Tris gels.

It is recommended for separating small- to medium-sized proteins. MES has a lower pKa than MOPS, making the MES SDS Running Buffer faster than the MOPS SDS running buffers. The difference in ion migration affects stacking and leads to a difference in protein separation range between these buffers. The use of MOPS buffer allows proteins to run slower than when using MES buffer.

Med Biosciences MES-SDS running buffer provides higher resolution of small size protein bands, faster electrophoresis time, and better detection of lower amounts of protein on Bis-Tris gels than other standard electrophoresis buffers.

The 20X concentrated MES-SDS running buffer is composed of 1 M MES (2-[N-morpholino]ethanesulfonic acid), 1 M Bis-Tris, 20 mM EDTA, 2% SDS, and other ingredients (pH 7.3 ± 0.1).

The 4X LDS Sample Loading Buffer contains 10% Glycerol, a mixture of Tris-Base and Tris-HCL (pH 8.5), Lithium Dodecyl Sulfate (LDS), EDTA, and dye mixture.

The protein reducing agent DTT is provided in a powder form. Add 50 µl or 100 µl deionized water to DTT vial to get a final concentration of 500 mM (10X). Mix well to dissolve and store it at – 20°C after use.

Antioxidant prevents sample reoxidation and maintains proteins in a reduced state during protein gel electrophoresis and protein transfer. It is added to the running buffer when performing protein gel electrophoresis under reducing conditions. Our Protein Antioxidant migrates with reduced proteins to maintain reducing conditions and to prevent reoxidation of sensitive amino acids such as methionine and tryptophan. The antioxidant is provided in 200X concentrate and is composed of 10% N,N-dimethylformamide.

PROCEDURE:

Add LDS sample buffer (4X) to the protein sample, maximum 40 µg, (with or without reducing agent, 1/10 of the final volume), and heat at 70 ᴼC for 10 min. Centrifuge the heated samples at 2,400 × g for 30 s to bring down insoluble material.

Dilute MES SDS running buffer (20X) to a 1X solution using ultrapure water. Make up to 1 L.

Add 1 volume of the antioxidant solution (200X) to 199 volumes of the 1X MES-SDS Buffer. Fill the electrophoresis tank with this buffer. Note: antioxidant is only used with reducing SDS-PAGE, i.e., with samples incubated with LDS loading buffer containing DTT.

Load the prepared heated samples on the Bis-Tris gel and run at 180 V constant voltage and 35 min run time, i.e., until the bromophenol blue tracking dye elutes from the gel at approximately 30–35 min.

PACKAGING:

Available in 2 kit sizes as shown in the above table.

Minimum order: 1 Kit

We accommodate customized packaging requests. Please contact us with your request for customized packaging.

STORAGE TEMP

The kit is shipped at ambient temperature. Store at 4 ᴼC.

SHELF LIFE

12 Months

NOTES

Molecular/Electrophoresis grade buffer, filter sterile, and DNase/RNase and protease free.

Each batch of buffers undergoes strict quality control and standard operating procedures to ensure high quality and lot-to-lot consistency.

Minimum order: 1 Kit.

Request a discounted quote for quantities ≥ 3 Kits.

This product is developed, designed, and sold exclusively for research purposes and in vitro use only.