What is Perm buffer?
Tonbo Biosciences Flow Cytometry Perm Buffer (10X) is provided as a concentrate which, when diluted with distilled water to a 1X solution, provides best results during intracellular staining of cytokines and other cytoplasmic antigens, by maintaining membrane permeabilization throughout staining and washing steps.
What is stain buffer?
Cell Staining Buffer is an antibody diluent and cell wash buffer optimized for use in immunofluorescent staining of viable or fixed single cell suspensions. Cell Staining Buffer contains bovine calf serum as a protein carrier to reduce non-specific binding of antibodies and fluorochrome reagents to target cells.
What is BSA staining?
Description. PharmingenStain Buffer (BSA) can be used for the immunofluorescent staining of single-cell suspensions prepared from either lymphoid tissues, bone marrow, peripheral blood, or cultured cells.
How do you use GolgiStop?
Procedure for Using BD GolgiStop™: Add 4 µl of BD GolgiStop™ for every 6 mL of cell culture (e.g., ~10^6 cells/mL) and mix thoroughly. Treatment of stimulated cells for 4 to 6 hours with BD GolgiStop™ significantly increases the ability to detect cytokine-producing cells by immunofluorescent staining.
How do you prepare a permeabilization buffer?
1X Phosphate Buffered Saline (PBS): To prepare 1 L 1X PBS: add 100 ml 10X PBS (#12528) to 900 ml water, mix. Cell Permeabilization Buffer: Purchase ready-to-use (#39487) or to prepare 10 ml, add 30 µl Triton™ X-100 to 10 ml Antibody Dilution Buffer. Store at 4°C.
What is cell staining?
Cell staining is a technique that can be used to better visualize cells and cell components under a microscope. By using different stains, one can preferentially stain certain cell components, such as a nucleus or a cell wall, or the entire cell.
What is BSA buffer?
BSA (bovine serum albumin) is a blocking buffer for IHC, ICC, ELISA and Western blotting; used to block non-specific antibody binding. Commonly used in cell culture. Purity: ≥98%
Why is BSA used for blocking?
BSA blocking is a routine practice among clinicians and researchers working on immunoassays throughout the world. The primary role of BSA is to prevent the non-specific binding by blocking the leftover spaces over solid surface after immobilization of a capture biomolecule.
What does Golgi plug do?
BD GolgiStop (monensin) works by accumulation of protein at the endoplasmic reticulum (ER) stage, while BD GolgiPlug (brefeldin) works with accumulation of protein at the golgi complex. The use of these inhibitors leads to an enhanced ability to detect cytokine-producing cells by intracellular flow cytometry.
How does PMA ionomycin work?
PMA activates protein kinase C, while ionomycin is a calcium ionophore, and stimulation with these compounds bypasses the T cell membrane receptor complex and will lead to activation of several intracellular signaling pathways, resulting in T cell activation and production of a variety of cytokines.
Why do you have to permeabilize cells?
The permeabilization step removes more cellular membrane lipids to allow large molecules like antibodies to get inside the cell.
What is cytoperm solution used for?
Description BD Cytofix/Cytoperm™ solution can be used for the simultaneous fixation and permeabilization of cells prior to intracellular cytokine staining. Preparation and Storage Store undiluted at 4° C. Cytofix/Cytoperm solution is supplied as a 1X solution.
What is the function of a buffer?
Buffers are used to maintain a stable pH in a solution, as they can neutralize small quantities of additional acid of base. For a given buffer solution, there is a working pH range and a set amount of acid or base that can be neutralized before the pH will change.
Is cytoperm harmful to humans?
Warnings and Precautions:BD Cytofix/Cytoperm contains formaldehyde, a suspected carcinogen. Avoid contact with skin, eyes, and mucous membranes. Avoid breathing fumes. Harmful by inhalation, in contact with skin, and if swallowed.
What is the size of BD cytoperm solution?
Material Number: 554722 Size: 125 ml Description BD Cytofix/Cytoperm™ solution can be used for the simultaneous fixation and permeabilization of cells prior to intracellular cytokine staining. Preparation and Storage Store undiluted at 4° C. Cytofix/Cytoperm solution is supplied as a 1X solution. Application Notes