- Detergents for Cell Lysis and Protein Extraction
- GDN - Digitonin's synthetic alternative
- GDN or DSP: How to decide
- GDN101 - GDN
- GDN - Digitonin's synthetic alternative
Detergents for Cell Lysis and Protein ExtractionCreate mode — the default mode when you create a requisition and PunchOut to Bio-Rad. You can create and edit multiple shopping carts. Edit mode — allows you to edit or modify an existing requisition prior to submitting. You will be able to modify only the cart that you have PunchedOut to, and won't have access to any other carts. Inspect mode — when you PunchOut to Bio-Rad from a previously created requisition but without initiating an Edit session, you will be in this mode. You cannot modify any Cart contents. Click here to find out how. Download PDF. The detection of intracellular antigens requires a cell permeabilization step prior to staining. This method provides an alternative procedure for use when protocol FC7. In some cases specific recommendations are provided on product datasheets, and these methods should always be used in conjunction with the product and batch specific information provided with each vial. A certain level of technical skill and immunological knowledge is required for the successful design and implementation of these techniques; these are guidelines only and may need to be adjusted for particular applications. Note: This procedure causes a reduction in Forward Scatter FSC signal, so the flow cytometer set up may need to be adjusted to compensate. You can create and edit multiple shopping carts Edit mode — allows you to edit or modify an existing requisition prior to submitting. You will be able to modify only the cart that you have PunchedOut to, and won't have access to any other carts Inspect mode — when you PunchOut to Bio-Rad from a previously created requisition but without initiating an Edit session, you will be in this mode. Third-Party Cookies being blocked. Please Note. Direct immunofluorescence staining of intracellular antigens: Digitonin method. If required perform staining of cell surface antigens using appropriate directly conjugated monoclonal antibodies at 4 o C, avoiding direct light. Discard supernatant. Incubate for 20 minutes at room temperature. Wash twice with 3ml 0. Pellet cells at g and 4 o C for 5 min. Decant supernatant. Wash twice with 3 ml 0. After each wash step pellet cells at g and 4 o C for 5 minutes.
GDN - Digitonin's synthetic alternative
Need a reason to switch from Digitonin to GDN? How about a whole grid box full? Extracted from the foxglove plant, Digitalis purpureaDigitonin has been widely used for the solubilization and purification of membrane proteins. In recent years, Digitonin has been used in the Cryo-EM structure determination of membrane proteins, being used in the determination of 6 of the 33 unique membrane protein structures determined by this method. Being a natural product, there are several drawbacks to Digitonin, including batch to batch variability, low solubility, high toxicity, and price. First characterized inGDN contains a steroid-based hydrophobic group linked to a di-maltose head group Fig. Being a synthetic compound, there is no batch-to-batch variability, and you don't have to worry about harmful toxic byproducts commonly found in Digitonin such as digitoxin, digoxin, and other cardiac and steroidal glycosides. Figure 1: Structure of GDN. To build on the excitement from these new structures, we are happy to offer a new 25 gram pack size of GDN, which offers significant savings over our smaller pack sizes. Have a GDN success story to share? Let us know and we'll be happy to feature your work in an upcoming newsletter! Magnani, F. Laguerre, A. Chae, P. Zhang, Y. Guo, H. Saotome, K.
GDN or DSP: How to decide
GDN101 - GDN
Structure of surfactants. Generalized structure of a single detergent molecule top and the complete structure of CHAPS bottoman example of a zwitterionic detergent. Detergents are amphipathic molecules, meaning they contain both a nonpolar "tail" having aliphatic or aromatic character and a polar "head". Ionic character of the polar head group forms the basis for broad classification of detergents; they may be ionic charged, either anionic or cationicnonionic unchargedor zwitterionic having both positively and negatively charged groups but with a net charge of zero. Like the components of biological membranes, detergents have hydrophobic-associating properties as a result of their nonpolar tail groups. Nevertheless, detergents are themselves water-soluble. Consequently, detergent molecules allow the dispersion miscibility of water-insoluble, hydrophobic compounds into aqueous media, including the extraction and solubilization of membrane proteins. Detergents at low concentration in aqueous solution form a monolayer at the air—liquid interface. At higher concentrations, detergent monomers aggregate into structures called micelles. A micelle is a thermodynamically stable colloidal aggregate of detergent monomers wherein the nonpolar ends are sequestered inward, avoiding exposure to water, and the polar ends are oriented outward in contact with the water. Both the number of detergent monomers per micelle aggregation number and the range of detergent concentration above which micelles form called the critical micelle concentration, CMC are properties specific to each particular detergent see table. The critical micelle temperature CMT is the lowest temperature at which micelles can form. The CMT corresponds to what is known as the cloud point since detergent micelles form crystalline suspensions at temperatures below the CMT and are clear again at temperatures above the CMT. Detergent properties are affected by experimental conditions such as concentration, temperature, buffer pH and ionic strength, and the presence of various additives. For example, the CMC of certain nonionic detergents decreases with increasing temperature, while the CMC of ionic detergents decreases with addition of counter ion as a result of reduced electrostatic repulsion among the charged head groups. In other cases, additives such as urea effectively disrupt water structure and cause a decrease in detergent CMC. Generally, dramatic increases in aggregation number occur with increasing ionic strength. Detergents can be denaturing or non-denaturing with respect to protein structure. Denaturing detergents can be anionic such as sodium dodecyl sulfate SDS or cationic such as ethyl trimethyl ammonium bromide. These detergents totally disrupt membranes and denature proteins by breaking protein—protein interactions. Non-denaturing detergents can be divided into nonionic detergents such as Triton X, bile salts such as cholate, and zwitterionic detergents such as CHAPS. Although detergents are available from several commercial sources and used routinely in many research laboratories, the importance of detergent purity and stability is not widely appreciated. Detergents often contain trace impurities from their manufacture. Some of these impurities, especially peroxides that are found in most nonionic detergents, will destroy protein activity. In addition, several types of detergents oxidize readily when exposed to the air or UV light, causing them to lose their properties and potency as solubilizing agents. We offer several high purity, low peroxide—containing detergents that are packaged under nitrogen gas in clear glass ampules. These Thermo Scientific Surfact-Amps Detergent Solutions provide unsurpassed convenience, quality and consistency for all detergent applications. A sampler kit includes 10 different purified detergents seven in the Surfact-Amps format and three in solid form. A major factor determining the behavior and interaction of molecules in biological samples is their hydrophilicity or hydrophobicity.