Lab Tools

COATING STABILIZER & BLOCK BUF

Insulin-Transferrin-Selenium-Ethanolamine (ITSE) is a basal media supplement and can be used to reduce the amount of fetal bovine serum (FBS) needed to culture cells. Insulin promotes the uptake of glucose and amino acids, transferrin is a non-toxic iron carrier, selenium is required for the function of glutathione peroxidase and other antioxidant enzymes, and ethanolamine is a phospholipid precursor. The components of the solution are manufactured animal-free and it is provided as a 100x sterile concentrate, 10mL of ITSE concentrate is sufficient for 1 liter of medium.

Insulin-Transferrin-Selenium (ITS) is a basal media supplement and can be used to reduce the amount of fetal bovine serum (FBS) needed to culture cells. Insulin promotes the uptake of glucose and amino acids, transferrin is a non-toxic iron carrier, selenium is required for the function of glutathione peroxidase and other antioxidant enzymes. The components of the solution are manufactured animal-free and it is provided as a 100x sterile concentrate, 10mL of ITS concentrate is sufficient for 1 liter of medium.

BrdU is a synthetic nucleoside that is an analog of thymidine and is commonly used in the detection of proliferating cells in living tissues. It is reported that BrdU can be used to promote chemical induction of mouse iPSCSs in conjunction with other chemical reagents including, CHIR 99021, ALK5 Inhibitor (RepSox), and Forskolin. BrdU is also reported to induce a sustained reduction in the proliferation rate of cancer cells.

CFSE is a reagent useful for cell tracking, proliferation studies, and cell motility studies. After passively diffusing into cells, it is converted to a fluorescent carboxyfluorescin molecule as its acetate groups are cleaved by intracellular esterases. The fluorescent CFSE is retained inside the cell for extended periods, because the molecules forms stable covalent crosslinks with the intracellular proteins. At each cell division, the intensity of the CFSE fluorescence is halved. The peak excitation of the molecule is 494 nm and the peak emission is 521 nm and it may be used with standard fixatives containing formaldehyde and permeabilization buffers that contain saponin.

The Cell Stimulation and Protein Transport Inhibitor Cocktail is a pre-mixed cocktail used to induce cytokine production and inhibit their intracellular transport for flow cytometry detection. The solution is composed of an optimized mixture of brefeldin A, ionomycin, monensin, and PMA. PMA and ionomycin induce cytokine production in many cell types and brefeldin A and monensin are protein transport inhibitors.

Red Blood Cell Lysing Buffer, containing ammonium chloride, is formulated to lyse RBC from human and murine samples while leaving the lymphocytes intact. This buffer, supplied at 10X dilution, should be diluted to 1X to produce the desired effect.

The permeabilization buffer can be used for intracellular staining of cytoplasmic and nuclear antigens. The buffer maintains cell permeability throughout the staining process. It has been formulated to reduce background non-specific staining and increase the specific fluorescence signal. The 10x concentrate solution should be diluted with distilled water to 1x prior to use.

The Transcription Factor Fixation/Permeabilization solution can be used with the relevant antibodies to stain for transcription factors such as Foxp3 and intranuclear staining of cytokines, chemokines, and nuclear proteins. The concentrated solution should be used with the Transcription Factor Fixation/Permeabilization Diluent (cat#92555-00) and Permeabilization Buffer (cat# 92110-00).