Learn More
Invitrogen™ Alexa Fluor™ 555 Hydrazide
Description
Alexa Fluor™ 555 Hydrazide is useful as a cell tracer and as a reactive dye for labeling aldehydes or ketones in polysaccharides or glycoproteins. Alexa Fluor™ 555 is a bright, orange fluorescent dye. Used for stable signal generation in imaging and flow cytometry, Alexa Fluor™ 555 dye is water soluble and pH-insensitive from pH 4 to pH 10. In addition to reactive dye formulations, we offer Alexa Fluor™ 555 dye conjugated to a variety of antibodies, peptides, proteins, tracers, and amplification substrates optimized for cellular labeling and detection.
Detailed information about this AlexaFluor™ hydrazide:
• Fluorophore label : Alexa Fluor™ 555 dye
• Reactive group: hydrazide
• Reactivity: Aldehydes or keytones in polysaccharides or glycoproteins
• Ex/Em of the conjugate: 554/567 nm
• Extinction coefficient: 159,000 cm-1M-1
• Spectrally similar dyes: TRITC, Cy3
• Molecular weight: ∼1,150
Cell Tracking and Tracing Applications
Alexa Fluor™ hydrazides and hydroxlamines are useful as low molecular weight, membrane-impermeant, aldehyde-fixable cell tracers, exhibiting brighter fluorescence and greater photostability than cell tracers derived from other spectrally similar fluorophores. They are easily loaded into cells by microinjection, infusion from patch pipette, or uptake induced by our Influx™ Pinocytic Cell-Loading Reagent (Cat. No. I14402).
Glycoprotein and Polysaccharide Labeling Applications
The Alexa Fluor™ hydrazides and hydroxlamines are reactive molecules that can be used to add a fluorescent label to biomolecules containing aldehydes or ketones. Aldehydes and ketones can be introduced into polysaccharides and glycoproteins by periodate-mediated oxidation of vicinal diols. Galactose oxidase can also be used to oxidize terminal galactose residues of glycoproteins to aldehydes.
Hydrazide vs Hydroxylamine
Hydrazine derivatives react with ketones and aldehydes to yield relatively stable hydrazones. Hydroxylamine derivatives (aminooxy compounds) react with aldehydes and ketones to yield oximes. Oximes are superior to hydrazones with respect to hydrolytic stability. Both hydrazones and oximes can be reduced with sodium borohydride (NaBH4) to further increase the stability of the linkage.
Learn More About Protein and Antibody Labeling
We offer a wide selection of Molecular Probes™ antibody and protein labeling kits to fit your starting material and your experimental setup. See our Antibody Labeling kits or use our Labeling Chemistry Selection Tool for other choices. To learn more about our labeling kits, read Kits for Labeling Proteins and Nucleic Acids—Section 1.2 in The Molecular Probes™ Handbook.
We’ll Make a Custom Conjugate for You
If you can’t find what you’re looking for in our online catalog, we’ll prepare a custom antibody or protein conjugate for you. Our custom conjugation service is efficient and confidential, and we stand by the quality of our work. We are ISO 13485:2000 certified.
Related Products
DMSO (dimethylsulfoxide) (Cat. No. D12345)
Antibody Conjugate Purification Kit for 0.5-1 mg (Cat. No. A33086)
Antibody Conjugate Purification Kit for 20-50 μg (Cat. No. A33087)
Antibody Conjugate Purification kit for 50-100 μg (Cat. No. A33088)
Specifications
Specifications
| Quantity | 1 mg |
| Product Type | Hydrazide |
| Content And Storage | Store at room temperature and protect from light. |
| Chemical Reactivity | Carboxylic Acid, Ketone, Aldehyde |
| Shipping Condition | Room Temperature |
| Reactive Moiety | Amine, Hydrazide |
| Label or Dye | Alexa Fluor™ 555 |
| Label Type | Alexa Fluor |
| Product Line | Alexa Fluor |
| Emission | 567 nm |
| Show More |
Frequently Asked Questions (FAQs)
The non-reactive Alexa Fluor and Alexa Fluor hydrazide derivatives may be used for injection site visualization. Other options include the fluorescent polystyrene microspheres, FluoSpheres, and dye-conjugated dextrans. The hydrazide derivatives and 'fixable' dextrans are retained by cross-linking using an aldehyde-based fixative.
Confirm that the tracer you are using crosslinks to proteins or has a primary amine for fixation-either a hydrazide, lysine fixable dextran, or a protein conjugate.
Use aldehyde-based fixatives to cross link the amines on the tracer.
Inject a larger amount or higher concentration of the tracer. Tracers are generally injected at 1-20% concentrations (10 mg/mL or higher).
Confirm that you are using the correct fluorescent filter for detection. You can perform a spot test by pipetting a small amount of the undiluted stock solution of the tracer onto a slide, then view under the filter you are using on your microscope. This will confirm if the tracer fluorescence can be detected and the fluorescent microscope filter is working properly.
Review tissue fixation and handling procedures to confirm if any reagents or processing procedures could be affecting the tracer.
Observing both types of transport is typical for biocytin. The conjugated cholera toxin subunit B products have been observed to travel only retrogradely.
Lucifer Yellow CH is a hydrazide, so any of our Alexa Fluor or fluorescent hydrazides could potentially be used. A listing of them can be found here. (https://www.thermofisher.com/us/en/home/life-science/cell-analysis/cell-tracing-tracking-and-morphology/neuronal-tracing/hydrazides-biocytins.html#prd)
Factors to consider are size of tracer, method of delivery (injection, direct application to tissue, etc.), and if the tracer needs to be fixable. Here are some links to details about the various classes of neuronal tracers we offer and how to choose between them:
Neuronal Tracing (https://www.thermofisher.com/us/en/home/life-science/cell-analysis/cell-tracing-tracking-and-morphology/neuronal-tracing.html)
Choosing a Tracer (https://www.thermofisher.com/us/en/home/references/molecular-probes-the-handbook/fluorescent-tracers-of-cell-morphology-and-fluid-flow/choosing-a-tracer.html)
Imaging Analysis (http://assets.thermofisher.com/TFS-Assets/BID/Reference-Materials/bioprobes-50-journal.pdf)
For Research Use Only. Not for use in diagnostic procedures.