Alexa Fluor® secondary antibodies

Alexa Fluor® conjugated secondaries are the brightest fluorescent secondary antibodies, created from Alexa Fluor® dyes. We offer secondary antibodies conjugated to nine Alexa Fluor® dyes that cover the whole spectrum with minimal spectral overlap, thus ideal for multi-color analysis and use in cell imaging and flow cytometry experiments.

Our range spans over 200 different Alexa Fluor® conjugated secondary antibodies, raised across many species, including donkey, goat, and rabbit. They can be used against specific IgG subclasses (eg, IgG2a and IgG2b) to detect multiple targets to which primary antibodies raised in different species are unavailable. To further minimize background from cross-species reactivity, we recommend using Alexa Fluor® pre-adsorbed secondary antibodies.

All products have been extensively tested within our facilities in applications such as cell imaging, flow cytometry, and fluorescent western blot with robust QC procedures.

Alexa Fluor® 488 secondary antibodies

Showing as yellow in the vial, Alexa Fluor® 488 is a green dye that matches the fluorescence of Cy2 and FITC. Excitation is ideally suited to 496 nm for argon ion lasers, with maximum absorption of 495 nm and maximum emission of 519 nm. The quantum yield is usually 0.92.

The ideal emission filter for 488 in confocal microscopy is either FITC or Alexa Fluor® 488, and for flow cytometry is 525/30 BP. Alexa Fluor® 488 is most compatible with DAPI, Alexa Fluor® 594, and Alexa Fluor® 647 for multiple staining.

Alexa Fluor® 647 secondary antibodies

Alexa Fluor® 647 is a far-red dye, showing blue in the vial, matching the fluorescence of APC and Cy5. This dye has a maximum absorption of 650 nm, a maximum emission of 665nm, and a quantum yield of around 0.33.

Alexa Fluor® 594 secondary antibodies

Just along the fluorescence spectrum from 568, Alexa Fluor® 594 is also a red dye, showing as purple in the vial, matching Texas Red in fluorescence. This dye exhibits a maximum absorption at 590 nm, a maximum emission at 617 nm, and a quantum yield of approximately 0.66.

Alexa Fluor® secondary antibodies leverage some of the brightest and most photostable dyes available, ensuring researchers can achieve clear, sharp imaging results. These antibodies are crucial for experiments requiring precise and multiple target detection within biological samples. Alexa Fluor® dyes, ranging from visible to near-infrared spectrum, enable simultaneous detection of multiple antigens, critical for complex cell imaging, flow cytometry, and fluorescent western blot applications.

We offer a comprehensive portfolio of Alexa Fluor® conjugated secondary antibodies, granting options suitable for a variety of species and IgG subclasses. This adaptability aids researchers when primary antibodies from different species are unavailable. To reduce cross-species reactivity and enhance specificity, we also supply pre-adsorbed options. Each batch of our Alexa Fluor® secondary antibodies undergoes stringent quality control processes. These include testing for intense staining capabilities and minimal background fluorescence, ensuring reliable and repeatable results.

To support rigorous research demands, our secondary antibodies are optimized for high dye-to-protein ratios, which minimize photobleaching and maximize fluorescence output across a range of pH conditions, maintaining stability and visibility in diverse experimental environments.