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S3401 Sigma

Sample Buffer, Laemmli 2× Concentrate

Synonym: SDS Sample Buffer

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Description

Other Notes

Solution contains 4% SDS, 20% glycerol, 10% 2-mercaptoethanol, 0.004% bromphenol blue and 0.125 M Tris HCl, pH approx. 6.8.

Quantity

Recommended usage: add 1 volume gel loading solution to 1 volume sample and mix well.

Application

Sample Buffer, Laemmli 2× concentrate has been used as a sample buffer for denaturing and loading of protein samples in SDS-PAGE.

Biochem/physiol Actions

Laemmli sample buffer is especially formulated for protein sample preparation to be used in the Laemmli SDS-PAGE system. A protein sample is mixed with the 2X sample buffer (1:1) and heated in boiling water for 2-5 min. The 2-mercaptoethanol reduces the intra and inter-molecular disulfide bonds. The SDS detergent denatures the proteins and subunits and gives each an overall negative charge so that each will separate based on size. The bromophenol blue serves as a dye front that runs ahead of the proteins and also serves to make it easier to see the sample during loading. The glycerol increases the density of the sample so that it will layer in the sample well.

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Amicon Pro Purification System
Protocols & Articles

Articles

A functional assay for microRNA target identification and validation

MicroRNAs (miRNA) are a class of small RNA molecules that regulate numerous critical cellular processes and bind to partially complementary sequences resulting in down-regulation of their target gene...
Joop Gäken, Azim M. Mohamedali, Jie Jiang, Farooq Malik, Doris Stangl, Alexander E. Smith, Constantinos Chronis, Austin G. Kulasekararaj, Nicolas S. B. Thomas, Farzin farzaneh, Mahavash Tavassoli, and Ghulam J. Maufi
Nucleic Acids Research 2012, 1-9
Keywords: Cellular processes, Sequences, Western blot

Peer-Reviewed Papers
15

References

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