FAQ Dyes & Lables

FAQ Dyes & Lables

Everything you wanted to know about dyes, labels, and labeling.

General Questions

We have compiled several application examples in the products section on our web site. There, you can find information about proven experimental conditions, about suitable combinations of dyes. Also, learn to take advantage of the particular properties of our dyes. If you have specific questions please don’t hesitate to contact us (info@abberior.com).

 

All abberior dyes sold for use in STED microscopy can equally well be applied in confocal microscopy. In fact, a good STED dye also makes a good dye for confocal microscopy.

In addition to our regular products displayed on our webpage we can provide additional products as antibodies with different specificities. However, since this requires a special production additional costs for custom production have to be charged.

Many abberior dyes are already available as hydrazides. If you are interested in these dyes or a custom product, please send us your request and we will do our best to provide you with a suitable solution.

The products do not contain preservatives like azides or thimerosal.

Our dye coupled antibodies are shipped at room temperature. After shipment they should be stored at 4 °C for short time periods. For extended time periods we recommend to aliquot the dyes and to store them at -20 °C to -80 °C.

Repeated freeze-thaw cycles should be avoided !

The exact concentration of the antibodies has to be determined empirically for every type of sample.

However, we recommend to use a dilution of 1:50 up to 1:200 as a starting point.

Embedding media

For our CAGE dyes conventional embedding media as PBS, MOWIOL, abberior MOUNT and others may be used.

Sealing of cover slips mounted with abberior MOUNT embedding media can be either done with two-component silicone-glue Twinsil®. (Picodent, Wipperfürth, Germany) or clear nail polish (e.g.wet n wild Shine Nail Color Clear Nail Protector).

Sealing of cover slips mounted with TDE Mounting media has to be done with clear nail polish (e.g.wet n wild Shine Nail Color Clear Nail Protector). Two-component silicone-glue Twinsil®. (Picodent, Wipperfürth, Germany) frequently does not polymerize completely.

Please do not use colored nail polish or nail polish containing metallic particles.

abberior offers different embedding media.

abberior MOUNT embedding media are aqueous embedding media with a refractive index in the range of 1.38. They are very convenient in terms of application. However, due to evaporation of contained water and polymerisation of the embedding medium (for abberior MOUNT SOLID), the refractive index will change over time.

TDE Mounting media are 2,2′-thiodiethanol based embedding media with refractive indices in the range of 1.408 – 1.518. For many applications TDE Mounting media are superior to aqueous embedding media. Especially for demanding applications as 4Pi microscopy, 3D STED microscopy or isoSTED microscopy, because the refractive index will stay constant over time.

In STORM, dSTORM and GSDIM microscopy, often embedding media are used that have to be prepared directly before measurement.
One reason for this is that compounds contained in these embedding media are degrading over time (i.e. enzymes, MEA and others). To this end we are not able to supply those embedding media.

For our CAGE dyes, conventional embedding media as PBS, MOWIOL, abberior MOUNT and others may be used.

NHS ester

N-Hydroxysuccinimidyl (NHS) esters of the carboxylic acids (RCOOH) look like RCOONS (where NS is N-succinimidyl residue). They readily react with primary amines (R’NH2) forming hydrolytically stable amides RCONHR’ and N-hydroxysuccinimide.

N-Hydroxysuccinimidyl carbonates of the alcohols (RCH2OH) look like RCH2OCONS, where NS is N-succiimidyl residue (as above). They readily react with amines R’NH2 forming hydrolytically stable urethanes RCH2OCONHR’ and N-hydroxysuccinimide.

N-hydroxysuccinimidyl carbonates are expected to react with amines a bit slower, but their stability is typically higher that that of N-hydroxysuccinimidyl esters.

The stability of the NHS esters differs between the individual dyes.

For non-dissolved dyes which are kept at dry conditions at 4 °C the shelf life is typically 6 – 12 month. The shelf life increases significantly if they are stored at -20 °C or better -80 °C.

For dissolved NHS esters the shelf life depends strongly on the solvent used for dissolving or diluting the dye. Best use water free solvents.

In water containing solvents NHS esters are very labile.
Therefore we recommend dissolving NHS esters in water free dimethylformamide. Alternatively water free and very pure dimethylsulfoxide may be used.

Phalloidin

For labeling of mammalian cells a (methanol containing) stock solution of phalloidin needs to be diluted in aqueous buffer to a final concentration of 1 Unit/ ml. (Depending on the application, cell line etc.)

Phalloidin is shipped freeze-dried (i.e. in a solid form). Upon arrival it has to be dissolved in methanol (abs.). Depending on the dye the solubility of fluorophore coupled phalloidin is approx.. 0.1 – 0.5 mg/ ml (methanol).