What Is FITC And PE?

Can I use PE and PE cy7 together?

When using tandem antibody conjugates in multicolor staining panels, it is important to use exactly the same tandem conjugate for compensation tubes that are used for staining experiment samples.

PE-Cy7 (BD) ≠ PE-Cy7 (Biolegend) for any antibody conjugate..

What color is PE?

488 nm 532 nm 561 nm 575/26 PE (Exmax 496 nm/Emmax 578 nm) R-phycoerythrin (PE) is an accessory photosynthetic pigment found in red algae.

What does FITC mean?

Fluorescein IsothiocyanateFluorescein Isothiocyanate. Fluorescein isothiocyanate (FITC) is the most widely used fluorescent probe for the preparation of conjugates with biological molecules (Hansen, 1967; Haugland, 1990).

What type of Fluorochromes exist?

The most commonly used fluorophore is Fluorescein IsoThioCyanate (FITC). Today’s large selection of fluorophores consists of three groups: synthetic organic dyes (such as FITC), biological fluorophores such as the Green Fluorescent Protein (GFP), discussed below and Quantum Dots (QD) (see Chapter 4).

Can I use APC cy7 and PE cy7 together?

Most recent answer. In my experience, it is not advisable to use APC/Cy7 and PE/Cy7 together due to heavy cross-beam contamination as their emission max are exactly same. However, they can be used together if you are using machines like the Cytek Aurora.

What color is FITC?

green colorFITC has excitation and emission spectrum peak wavelengths of approximately 495 nm/519 nm, giving it a green color.

How do you choose Fluorochromes for flow cytometry?

Rule 1: Choose the brightest set of fluorochromes for your particular instrument configuration. Rule 2: Choose fluorochromes to minimize the potential for spectral overlap. Rule 3: Reserve the brightest fluorochromes for dim antibodies and vice versa.

How is flow cytometry compensation done?

How To Compensate A 4-Color Flow Cytometry Experiment Correctly4 Steps To Compensating A 4-Color Experiment. … Choose the correct carrier for compensation. … Step 2: Collect the data and make sure there is a sufficient number of events. … Calculate compensation correctly. … Apply the compensation values and inspect the results.

What makes a good fluorophore?

A fluorophore with good separation between the excitation and emission maxima typically results in more reliable detection than a fluorophore with little separation.

What is PE flow cytometry?

R-PE is a large molecule used for fluorescence-based detection, primarily in flow cytometry, microarray assays, ELISAs, and other applications that require high sensitivity but not photostability. … Long-wavelength fluorescence emission.

Is GFP a fluorophore?

The Chromophore of GFP. GFP is unique among fluorescent proteins in that its fluorophore is not a seperately synthesized prostethic group but composed of modified amino acid residues within the polypeptide chain.

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.

Why use isotype controls flow cytometry?

Isotype controls are primary antibodies that lack specificity to the target, but match the class and type of the primary antibody used in the application. Isotype controls are used as negative controls to help differentiate non-specific background signal from specific antibody signal.

Why flow compensation is required?

The purpose of steam flow meter with temperature and pressure compensation is for when the volume deviate the actual flow compensation will correct the result well. Steam in gas status, the density varies with the changes of temperature and pressure itself. If don’t have the compensation will cause a huge error.

Why is compensation of fluorescence necessary?

However, when emission spectra overlap, fluorescence from more than one fluorochrome may be detected. To correct for this spectral overlap, a process of fluorescence compensation is used. This ensures that the fluorescence detected in a particular detector derives from the fluorochrome that is being measured.