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Detection of Botulinum Neurotoxin Types C and D by Immunoaffinity Chromatography (CAT#: STEM-ACM-0102-CJ)

Introduction

Clostridium botulinum is a gram-positive, anaerobic, and spore-forming soil bacterium that can be found in the guts of domestic animals. The fact that it produces the most toxic metabolite known to humans brought it to the attention of medical microbiologists in the early days of this field. Nowadays, seven distinct botulinum neurotoxins (BoNTs) are known. In the order of their discovery, they have been named types A to G.




Principle

Immunoaffinity chromatography is a method for separating target antibodies or antigens from a heterogeneous solution. It is column-based, which means that the solution is flowed through a column and eluted at the other end. The column is pre-functionalized with the capture antibody or antigen. The target protein is adsorbed onto the resin-bound capture protein and is retained in the column while the remaining solution is eluted. The fraction containing the target protein is later eluted and purified.

Applications

Environmental Analysis; Microbiology

Procedure

1. Preparation of Column: The column is loaded with solid support such as sepharose, agarose, cellulose etc.. Ligand is selected according to the desired isolate. Spacer arm is attached between the ligand and solid support.
2. Loading of Sample: Solution containing a mixture of substances is poured into the elution column and allowed to run at a controlled rate.
3. Elution of Ligand-Molecule Complex: Target substance is recovered by changing conditions to favor elution of the bound molecules.

Materials

• Sample: Plants; Natural Food; Protein; Drug; Pollutants; Blood; Saliva; Serum; Plasma; Antibodies; Viruses & More
• Equipment: Agarose; Silica gel; Aluminium oxide; Acrylate; Organic polymers; Wash Buffer
• (Optional): Ligand; Spacer arm; Column

Notes

1. Immunoaffinity chromatography is one of the most widely used affinity-derived procedures, and it permits the production of ligands when the needed ligand is unavailable.
2. In this method, the stationary phase consists of an antibody or an agent linked to antibodies. Due to the high specificity of antibodies, it is possible to isolate varied substances using this method.
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