What is the wavelength of mCherry?
mCherry absorbs light between 540-590 nm and emits light in the range of 550-650 nm.
What is the difference between mCherry and GFP?
GFP is more sensitive to pH change than mCherry. The pH-dependent emission profile of GFP and mCherry was investigated by resuspending GFP and mCherry in phosphate buffers with a pH ranging from 4.4 to 9.1. The emission was then plotted against the pH value to obtain the profile.
What is mCherry fluorescence?
mCherry is a bright red monomeric fluorescent protein created by rounds of directed evolution of DsRed. mCherry matures rapidly, making it possible to see results very soon after transfection or activation of transcription. It is highly photostable and resistant to photobleaching (Shaner et al. 2004).
What channel is mCherry on?
Imaging
| Fluorescent protein | Excitation maximum (nm) | Cytometer channel |
|---|---|---|
| mOrange2 | 549 | PE |
| mApple | 568 | PE |
| tdTomato | 554 | PE |
| mCherry | 587 | PI |
What wavelength is mCherry excited at?
587 nm
mCherry has a single photon excitation peak at 587 nm and an emission peak at 610 nm [7].
Can I use mCherry and GFP?
Due to the spectral properties of GFP and mCherry, they are considered an ideal combination for co-localisation and co-expression experiments.
What wavelength is GFP?
Green fluorescent protein (GFP) is the most commonly used fluorescent reporter. Initially isolated from the jellyfish Aqueoria victoria, GFP has since been engineered to generate the brighter and more photostable enhanced GFP (eGFP), with an excitation wavelength of 488 nm and emission wavelength of 509 nm.
How do you detect mCherry?
It’ll depend on the flow cytometer you have access to but to best detect the mCherry signal excite with the yellow-green laser at 561 nm and detect in the PE-TexasRed channel with a 610/20 bandpass filter.
What laser excites mCherry?
561-nm laser
In addition, the 561-nm laser efficiently excites fruit fluorescent proteins such as mCherry.
What is the half life of mCherry?
about 40 minutes
mCherry has a maturation half-time of about 40 minutes, whereas sfGFP becomes fluorescent much faster, on the order of a few minutes.
What wavelength is DAPI?
358 nm
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). Therefore, for fluorescence microscopy, DAPI is excited with ultraviolet light and is detected through a blue/cyan filter.
Is GFP same as FITC?
Green Fluorescent Protein (GFP) Antibody (FITC) is a Rabbit Polyclonal antibody conjugated to FITC against GFP. IP: 1/50 – 1/200. Optimal dilutions/concentrations should be determined by the end user.
Is mCherry and RFP the same?
MCherry, the Discosoma sp. mushroom coral-derived monomeric red fluorescent protein (RFP), is a commonly used genetically encoded fluorophore for live cell fluorescence imaging.
What is the best red fluorescent protein?
mCherry is the most widely used and cited red fluorescent protein owing to its fast maturity, stability, and resistance to photobleaching. mCherry and DsRed-Monomer are ideal for tagging proteins with diverse functions and/or subcellular localization patterns.
What wavelength is RFP?
Red fluorescent protein (RFP) is a versatile biological marker for monitoring physiological processes, visualizing protein localization, and detecting transgenic expression in vivo. RFP can be excited by the 488 nm or 532 nm laser line and is optimally detected at 588 nm.
What is the difference between DAPI and Hoechst?
Hoechst dyes are typically used for staining DNA content in live cells due to its high cell membrane permeability. DAPI is typically used for staining DNA content in fixed cells due to its low membrane permeability.
What wavelength is Tritc?
544 nm
TRITC (Tetramethylrhodamine-isothiocyanate) is a fluorescent compound with an excitation peak at 544 nm and an emission peak at 570 nm.
What wavelength is FITC?
FITC emits fluorescence from 475 to 650 nm, peaking at 525 nm, which falls in the green spectrum.
At what wavelength does GFP fluorescence?
What is the excitation wavelength of DAPI?
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. It is excited by the violet (405 nm) laser line and is commonly used as a nuclear counterstain in fluorescence microscopy, flow cytometry, and chromosome staining.
What Colour is 488 wavelength?
green-fluorescent
Invitrogen Alexa Fluor 488 dye is a bright, green-fluorescent dye with excitation ideally suited to the 488 nm laser line.
Is Alexa Fluor 488 same as FITC?
Alexa Fluor 488 (Ex-Max 495 nm/Em-Max 519 nm): have nearly identical emission and excitation maxima as FITC. However, Alexa Flour 488 display higher photostability and is less susceptible to self-quenching.
Is GFP and FITC the same?
Green Fluorescent Protein (GFP) Antibody (FITC) is a Rabbit Polyclonal antibody conjugated to FITC against GFP.
What wavelength does GFP absorb at?
395 nm
A mechanism for the fluorophore formation has been proposed (3) but needs to be confirmed by further studies. GFP absorbs blue light at 395 nm, with a smaller peak at 475 nm, and emits green light at 508 nm with a quantum yield of 0.72–0.85 (12, 13).