orange-fluorescent Invitrogen Alexa Fluor 546 dye is a bright, orange-fluorescent dye that can be excited using the 488 nm or 532 nm laser lines. For stable signal generation in imaging and flow cytometry, Alexa Fluor 546 dye is pH-insensitive over a wide molar range.
How does Alexa Fluor work?
Alexa Fluor dyes can be conjugated directly to primary antibodies or to secondary antibodies to amplify signal and sensitivity or other biomolecules. The excitation and emission spectra of the Alexa Fluor series cover the visible spectrum and extend into the infrared.
What Alexa 532?
Alexa Fluor 532 is a yellow-fluorescent dye with excitation ideally suited for the frequency-doubled Nd:YAG laser line. Alexa Fluor 532 dye is pH-insensitive over a wide molar range, helping ensure that you get stable signals in live-cell imaging applications.
Is Alexa 555 red?
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 wavelength is DAPI?
358 nm 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.
How long do Alexa Fluor antibodies last?
Storage at 4°C should not exceed 1 or 2 weeks.
What does Alexa Fluor bind to?
Probes with high fluorescence quantum yield and high photostability allow detection of low-abundance biological structures with great sensitivity. Alexa Fluor 488 dye molecules can be attached to proteins at high molar ratios without significant self-quenching, enabling brighter conjugates and more sensitive detection.
Are Alexa Fluor dyes fixable?
All Alexa dyes and their conjugates are more fluorescent and more photostable than their commonly used spectral analogues listed above. … Alexa dye derivatives of phalloidin stain F-actin with high specificity. Hydrazide forms of the Alexa dyes are very bright, formaldehyde-fixable polar tracers.
What color is Cy3?
Cy3 dye dashboard The Cy3 dye is a traditional orange-fluorescent label for protein and nucleic acid conjugates used for imaging, flow cytometry, and genomic applications.
Is FITC same as 488?
High-performance alternatives to fluorescein The Invitrogen Molecular Probes Alexa Fluor 488 dye—with spectral properties and quantum yield nearly identical to those of fluorescein isothiocyanate (FITC)—produces brighter, more photostable conjugates.
What color is Alexa Fluor 633?
red Alexa Fluor® 633 is a bright and photostable far-red dye with excitation ideally suited to the 633 nm laser line. Used for stable signal generation in imaging and flow cytometry, Alexa Fluor® 633 dye is water soluble and pH-insensitive from pH 4 to pH 10.
What color is Alexa Fluor 555?
Orange Table 1. Properties of Alexa Fluor Dyes
| Alexa Fluor dye | Absorption max. (nm) | Emission color* |
|---|---|---|
| Alexa Fluor 555 | 555 | Orange |
| Alexa Fluor 568 | 578 | Orange/Red |
| Alexa Fluor 594 | 590 | Red |
| Alexa Fluor 610 | 612 | Red |
What Colour is Alexa Fluor 647?
red 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 is a FAM dye?
Fluorescein amidites, abbreviated as FAM, are important synthetic equivalents of fluorescein dye used in oligonucleotide synthesis and molecular biology. … Oligonucleotides labeled with fluorescein at one of the termini and with a quencher at the other can serve as molecular beacons.
Can you fix DAPI?
we had good success with DAPI-staining (included into the mounting medium) after fixation in 4% buffered formol. The good old neutral buffered formalin (NBF) for 30 mins is the recommended choice. The just wash with PBS and stain with 1:1000 dilution from a 1 mg/ml solution of DAPI in water.
How do I see DAPI?
First, fix and permeabilize cultured cells with a protocol appropriate for your sample.
- Wash the cells 1–3 times in PBS as needed.
- Add sufficient 300 nM DAPI stain solution to cover the cells.
- Incubate for 1–5 minutes, protected from light.
- Remove the stain solution.
- Wash the cells 2–3 times in PBS.
- Image the cells.
What is DAPI blue?
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.
Can I leave secondary antibody overnight?
Generally, maximum blocking time should not exceed 2 hours at room temperature or proteins can be exchanged from the membrane. … Incubate the membrane in the secondary antibody reagent of choice for 30 minutes to 1 hour at room temperature or overnight at 4°C with agitation.
Do you wash after secondary antibody?
After incubating with the secondary antibody, the membrane is then washed with TBST or PBST buffer for 4-5 times, about 5 minutes each time with agitation, to remove residual unbound secondary antibody.
Can secondary antibodies be reused?
Finally, very few researchers will reuse secondary antibodies, as you generally burn through fewer of them per equivalent volume. Secondary antibodies are also cheaper and used at far more dilute ranges.
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.
Which Alexa Fluor is 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.
Is af647 same as APC?
5 and APC-Cy7. 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.
What is DAPI staining used for?
DAPI staining was used to determine the number of nuclei and to assess gross cell morphology. Following light microscopic analyses, the stained cells were processed for electron microscopy.
Why is DyLight 488 Essential?
DyLight® 488 is a green fluorophore commonly used in applications such as protein labeling, immunolabeling, fluorescence microscopy, and enzyme immunoassays. … DyLight® 488 is bright and photostable, and can be used as a substitute for other green fluorophores such as FITC.
How do you dissolve Alexa Fluor?
The Alexa Fluor® NHS ester is typically dissolved in high-quality anhydrous dimethylformamide (DMF) or dimethylsulfoxide (DMSO) (D12345), and the reaction is carried out in 0.1–0.2 M sodium bicarbonate buffer, pH 8.3, at room temperature for 1 hour.
What does Texas red stain?
Texas Red or sulforhodamine 101 acid chloride is a red fluorescent dye, used in histology for staining cell specimens, for sorting cells with fluorescent-activated cell sorting machines, in fluorescence microscopy applications, and in immunohistochemistry.
What Cy3 and Cy5 denotes in microarray technology?
The microarray is analyzed in a scanner, which uses laser light to cause each spot to fluoresce and measures the relative intensity of emission, typically at green (Cy3) and red (Cy5) wavelengths.
What is Cyanine Blue?
1a : a strong blue that is greener and deeper than Sèvres and redder and darker than cerulean blue (see cerulean blue sense 1b) — called also Leitch’s blue.
Graduated from ENSAT (national agronomic school of Toulouse) in plant sciences in 2018, I pursued a CIFRE doctorate under contract with Sun’Agri and INRAE in Avignon between 2019 and 2022. My thesis aimed to study dynamic agrivoltaic systems, in my case in arboriculture. I love to write and share science related Stuff Here on my Website. I am currently continuing at Sun’Agri as an R&D engineer.