Wednesday, January 30, 2013
Protein Quantification (Bradford Assay)
Bradford assay is a rapid and accurate method to determine the concentration of protein. The assay is based on the observation that the absorbance maximum for an acidic solution of Coomassie Brilliant Blue G-250 shifts from 465 nm to 595 nm when binding to protein occurs. Both hydrophobic and ionic interactions stabilize the anionic form of the dye, causing a visible color change. Within the linear range of the assay (~5-25 mcg/ml), the more protein present, the more Coomassie binds.
Bradford Assay
Experiment using Bradford Assay to determine protein concentration. Follows a specific protocol but can be used to understand how to perform a Bradford Assay.
How to Make an SDS-PAGE gel
Alright so here's a quick video on how to cast an SDS-PAGE gel. Although recipes can vary, the ingredients shown here are almost always used. Remember: always add TEMED last, and pour into plates immediately after!
Already know how to make a gel, but have problems with leaky gels? Check out our other video on How to Avoid a Leaky SDS-PAGE gel: See the video here
Thursday, December 13, 2012
AFFINITY CHROMATOGRAPHY
It is mainly based on the biological affinity (or) biological specificity. This technique mainly requires previous knowledge of the molecule to be separated a specific ligand will only attach with a specific molecule.
Affinity Chromatography video here
