What is the best excitation light for FITC?

What is the best excitation light for FITC?

A fluorophore’s maximal absorbance informs you which laser line is optimal to be used for excitation. In the case of FITC, its maximum absorbance falls within the blue spectrum. Therefore, the blue 488 nm laser, which is close to FITC’s absorbance peak of 490 nm, is commonly used to excite this fluorophore.

Are FITC and GFP the same?

Green Fluorescent Protein (GFP) Antibody (FITC) is a Rabbit Polyclonal antibody conjugated to FITC against GFP.

What is the excitation for GFP?

GFP can be excited by the 488 nm laser line and is optimally detected at 510 nm.

What is GFP excitation wavelength?

The GFP from A. victoria has a major excitation peak at a wavelength of 395 nm and a minor one at 475 nm. Its emission peak is at 509 nm, which is in the lower green portion of the visible spectrum.

Is af488 brighter than FITC?

In addition, Alexa Flour 488 is pH insensitive (over a broad pH range) and tends to be brighter compared to FITC on most instruments.

Is FITC a bright fluorophore?

Fluorescein is a bright green fluorphore that is sensitive to changes in pH and can photobleach rapidly unless an antifade agent is used.

Which is better for FITC, GFP or FITC?

This is better for use of FITC filtersets as this mutant GFP has the same excitation range as FITC. Furthermore, the main laserline used for FITC excitation is from the argon laser at 488nm.

What is the excitation peak of a GFP?

The wild type like variants have their primary excitation peak centered on 395 nm, with an emission peak at 509 nm while the blue emitting mutants generally have an excitation peak at around 380 nm and an emission peak near 460 nm (Figure 1). Table 1. Excitation and emission data of GFP variants

What’s the maximum emission of a FITC molecule?

Figure 1 below illustrates the excitation and emission spectra of FITC under conditions of 488 nm excitation. FITC’s emission maximum is around 530 nm. This means that if we excite this molecule, there is a very high probability that it will emit green photons.

What kind of excitation filter is used for eGFP?

Fluorescence was determined using either a 485/20 excitation, 530/25 emission filter set or a 400/30 excitation, 508/20 emission filter set. For EGFP and wtGFP determinations using either filter set, a sensitivity setting of 170 was used.