What is Alexa Fluor 647?
Invitrogen Alexa Fluor 647 dye is a bright, far-red–fluorescent dye with excitation ideally suited for the 594 nm or 633 nm laser lines. For stable signal generation in imaging and flow cytometry, Alexa Fluor 647 dye is pH-insensitive over a wide molar range.
What wavelength is 647?
Alexa Fluor® 647 is a far-red fluorophore commonly used in applications such as immunolabeling, fluorescence microscopy, and flow cytometry. The dye has an absorption wavelength that peaks around 650 nm, and an emission maximum around 665 nm. Alexa Fluor® 647 is bright and photostable.
Is Alexa 647 Cy5?
For stable signal generation, Alexa Fluor 647 dye is generally brighter than Cy5 dye in protein and nucleic acid conjugates. Allophycocyanin (APC) is another bright fluorophore that is spectrally equivalent to Cy5 dye and Alexa Fluor 647 dye, but it is used almost exclusively in flow cytometry.
Is Alexa Fluor 647 APC?
Alexa Fluor 647 (Ex-Max 650 nm/Em-Max 668 nm): conjugates are highly photostable and remain fluorescent over a broad pH range. The excitation and emission maxima are nearly identical to those of APC, albeit APC tends to be brighter. However, Alexa Fluor 647 is considered more optimal for intracellular applications.
What is Alexa Fluor used for?
Alexa Fluor dyes are frequently used as cell and tissue labels in fluorescence microscopy and cell biology. Alexa Fluor dyes can be conjugated directly to primary antibodies or to secondary antibodies to amplify signal and sensitivity or other biomolecules.
What is FITC in flow cytometry?
Fluorescein isothiocyanate (FITC) is a derivative of fluorescein used in wide-ranging applications including flow cytometry. First described in 1942, FITC is the original fluorescein molecule functionalized with an isothiocyanate reactive group (−N=C=S), replacing a hydrogen atom on the bottom ring of the structure.
Which Alexa Fluor is blue?
Fluor 350
Dyes and chemistry
| Colour† | Emit (nm) | |
|---|---|---|
| Alexa Fluor 350 | Blue | 442 |
| — 405 | Blue | 421 |
| — 430 | Green | 541 |
| — 488 | Green | 519 |
Can I use PerCP and PerCP Cy5 5 together?
PerCP-‐Cy5. 5 overlap surprisingly a lot with BV711, don’t use together. PerCP overlap a bit with APC and PeCy7. Pe-‐CF594 spill over a lot from and to Pe, don’t use together.
Which fluorophores are brightest?
It is concluded that cyanine 3.18 should be useful for multicolor fluorescence experiments and that it may be the brightest fluorophore available for single-color fluorescence immunocytochemistry.
Which is brighter PE or APC?
Compared to PE, APC is not as”bright”. This is due in part to its lower extinction coefficient and quantum yield. It is also because APC is red-shifted compared to PE. Thus, the light source used to maximally excite APC will contain less energy than the one used to maximally excite PE.
Can I use APC Cy7 and PE Cy7 together?
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 Alexa Fluor 594?
red
Invitrogen Alexa Fluor 594 dye is a bright, red-fluorescent dye that can be excited using the 561 nm or 594 nm laser lines.
What color is Alexa Fluor 555?
Alexa Fluor® 555 (AF555, Alexa 555) is a yellow-emitting synthetic fluorophore that can be excited by the 532 nm yellow laser and captured with a 575/36 nm bandpass filter.
What is FITC conjugated?
FITC is a small organic molecule, and is typically conjugated to proteins via primary amines (i.e., lysines). Usually, between 3 and 6 FITC molecules are conjugated to each antibody; higher conjugations can result in solubility problems as well as internal quenching (and reduced brightness).
Is FITC same as GFP?
Just because GFP, CFSE and FITC are detected in the same channel (FL1), it does not mean you can use them interchangeably when setting up your compensation. GFP, CFSE and FITC are unique flourochromes and have different spillover into other channels.
Can I use FITC and PE together?
FITC overlap a lot with Pe, don’t use them together on same cells. PerCP-‐Cy5. 5 overlap surprisingly a lot with BV711, don’t use together.
Can I use PE and PE Cy5 together?
Pe-‐CF594 spill over a lot from and to Pe, don’t use together. PeCy5 spill over a lot to APC (bad), Pe and PeCy7. Avoid these combinations if possible.
What is the brightest fluorescent protein?
mNeonGreen is also the brightest monomeric green or yellow fluorescent protein yet described to our knowledge. Its high quantum yield and extinction coefficient (Table 1) make it between 1.5 and 3 times as bright as most commonly used green and yellow fluorescent proteins.
Is PE the same as R PE?
Phycoerythrin (PE) is available in two forms: R-PE and B-PE, though R-PE has found more use in flow cytometry (because of this, the shorthand PE is often used interchangeably with R-PE). PE has a molecular weight of 240,000 Daltons, with an extinction coefficient of 1,960,000 cm-1 M-1 and a quantum yield of 0.84.
Can you use APC and PE together?
Are PE and R PE the same?
Does PE and APC overlap?
First, the excitation spectra of PE and APC show little overlap. APC is maximally excited by 615 -655 nm light, but APC is not excited by 488 nm light that maximally excites PE.
What color is Alexa Fluor 568?
orange-
Invitrogen Alexa Fluor 568 dye is a bright, orange-fluorescent dye with excitation ideally suited for the 568 nm laser line on the Ar-Kr mixed-gas laser.
What Colour is Alexa Fluor 488?
green-fluorescent
Invitrogen Alexa Fluor 488 dye is a bright, green-fluorescent dye with excitation ideally suited to the 488 nm laser line.
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.