What type of cell causes apoptosis?
What type of cell causes apoptosis?
Killer lymphocytes
How do you induce apoptosis in cells?
Chemical induction of apoptosis
- Set up your cells for treatment: inoculate adherent cells into 10 cm2 tissue culture dishes or suspension cells into T75 flasks at a concentration of ~106 cells/mL.
- Add cellular damaging agents at the recommended concentrations to induce apoptosis.
How does annexin V work?
Annexin A5 (or annexin V) is a cellular protein in the annexin group. In flow cytometry, annexin V is commonly used to detect apoptotic cells by its ability to bind to phosphatidylserine, a marker of apoptosis when it is on the outer leaflet of the plasma membrane.
Can you fix annexin V?
Yes, cells can be fixed with formaldehyde after staining. Because Annexin V staining is dependent on calcium, all buffers used for washing and fixation should contain 1.25 mM CaCl2. Fixation may increase the background signal from Ethidium Homodimer III.
What is an apoptosis assay?
An apoptosis assay detects and quantifies the cellular events associated with programmed cell death, including caspase activation, cell surface exposure of phosphatidylserine (PS) and DNA fragmentation.
Is annexin V an antibody?
Annexin V Antibody recognizes endogenous levels of total annexin V protein. This antibody is not predicted to cross-react with other annexin family members.
What is Annexin V staining?
Annexin V staining is a common method for detecting apoptotic cells. Thermo Fisher Scientific offers high-quality fluorescent annexin V conjugates as standalone reagents and in a variety of kits for use in flow cytometry and for imaging suspension cells.
What is annexin V FITC?
Annexin V-FITC Apoptosis Staining / Detection Kit ab14085 is used in a 10 min, one-step staining procedure to detect apoptosis by staining phosphatidylserine molecules which have translocated to the outside of the cell membrane.
What color is FITC?
green
What is FITC used for?
Fluorescein-5-isothiocyanate (FITC) is a fluorescence dye and belongs to the xanthene dyes. FITC is used for labeling of different biomolecules, e.g. immunoglobulins, lectins and other proteins, peptides, nucleic acids, nucleotides; oligo-and polysaccha- rides.
What color is propidium iodide?
red
What is the difference between DAPI and Hoechst?
Hoechst dyes are often used as substitutes for another nucleic acid stain called DAPI. Key differences between Hoechst dyes and DAPI are: Hoechst dyes are less toxic than DAPI, which ensures a higher viability of stained cells. The additional ethyl group of the Hoechst dyes renders them more cell-permeable.
How do you dissolve propidium iodide?
To make a stock solution from the solid form, dissolve PI (MW = 668.4) in deionized water (dH2O) at 1 mg/mL (1.5 mM) and store at 4°C, protected from light. When handled properly, solutions are stable for at least 6 months.
What does DAPI label?
DAPI (4′,6-diamidino-2-phenylindole) is a blue-fluorescent DNA stain that exhibits ~20-fold enhancement of fluorescence upon binding to AT regions of dsDNA. DAPI is generally used to stain fixed cells since the dye is cell impermeant, although the stain will enter live cells when used at higher concentrations.
Does DAPI kill cells?
In S. cerevisiae, DAPI and Hoechst preferentially stain dead cells with nuclear and cytoplasmic localization. In live yeast, Hoechst shows dim nuclear and cytoplasmic staining, while DAPI shows dim mitochondrial staining. The dyes can be used to stain yeast at 12-15 ug/mL in PBS.
Does DAPI stain dead cells?
DAPI (4′,6-diamino-2-phenylindole, dihydrochloride) is a fluorescent nucleic acid stain that binds to minor grove A-T rich regions of double-stranded DNA. It is essentially excluded from viable cells, but can penetrate cell membranes of dead or dying cells.
Is DAPI cytotoxic?
In experiments, DAPI has high cytotoxicity, which reinforces the reason to avoid using DAPI for live cell staining. DAPI is fairly non-toxic to humans if exposure occurs.
Can DAPI stain RNA?
Fluorescence properties When bound to double-stranded DNA, DAPI has an absorption maximum at a wavelength of 358 nm (ultraviolet) and its emission maximum is at 461 nm (blue). DAPI will also bind to RNA, though it is not as strongly fluorescent. Its emission shifts to around 500 nm when bound to RNA.