How much biotin is coupled to my protein? The HABA/Avidin Assay
Tuesday, October 17, 2017
Biotin, a 244 Dalton molecule, exhibits an extraordinary binding affinity for avidin and streptavidin (K a =10 15 M -1 ). This high binding affinity means that proteins, particularly antibodies, are routinely labeled with biotin. The labeled proteins can now be rapidly purified with av... ..
Sputum Processing, from viscous mucus to homogenous samples
Tuesday, October 10, 2017
Sputum Liquefaction: from viscous, heterogeneous mucus to equivalent, homogenous samples Sputum processing has one main purpose; to render the viscous, heterogeneous specimen to a homogenous state (evenly distributed suspension). This homogenous suspension will be dispensed into mu... ..
High Efficiency & Stability Protein CrossLinking with EDC & NHS
Tuesday, September 26, 2017
EDC is the most popular zero-length crosslinker for biochemical conjugations because it can efficiently form conjugates between two protein molecules, between a protein and a peptide, and between proteins and oligonucleotides, and with small molecules. ..
Role of Non-Detergent Sulfobetaines in Protein Purification
Tuesday, September 12, 2017
There are several contemporary methods for protein purification. Some techniques are very powerful, such as reverse phase chromatography, immunoadsorption, or affinity chromatography. However, these methods may ultimately denature protein and negatively impact the desired product. Likewise, recom... ..
Ammonium Sulfate Protein Precipitation: The key to Salting-Out
Wednesday, September 6, 2017
Salt precipitation can be a very powerful tool to purify proteins by precipitation. Ammonium sulfate is usually the salt of choice since it is cheap, very soluble in water, and is able to become much more hydrated (interacts with more water molecules) than almost any other ionic solvent. In pract... ..
Modifying Oligonucleotide 5'-Phosphates By EDC for Improved Coupling
Tuesday, August 29, 2017
The heterobifunctional, zero-length carbodiimide crosslinker EDC is a versatile tool that can be used with Imidazole to modify, label, immobilize or bioconjugate oligonucleotides, DNA and RNA at their 5’ phosphate groups for a variety of applications: ..
Optimize Elution Conditions in Affinity Chromatography to Antibodies
Tuesday, August 22, 2017
Immunoaffinity purification utilizes the unique high specificity of purified antibodies ( polyclonal and monoclonal) immobilized onto a solid matrix (porous agarose beads) to rapidly and selectively purify target analytes from a complex mixture. Optimal elution conditions allow for the efficient ... ..
Immunoprecipitations (IP) for Protein Purification & Interactions Studies
Tuesday, August 15, 2017
ImmunoPrecipitation (IP) is a very useful immunochemical technique for isolating and purifying target proteins out of a complex sample (lysate). In conjunction with western blotting and/or other assay techniques, IPs can be used to determine: the presence and quantity of a protein; molecular weig... ..
How to select the Right protease inhibitor
Tuesday, August 8, 2017
While proteases are essential in maintaining the proper function of living cells, researchers often do not welcome their presence in their protein samples. Why would they when the mere presence of these enzymes jeopardizes the integrity of their samples and puts them one step closer to imminent d... ..