Donkey Anti-Mouse IgG (H+L) Secondary Antibody, Alexa Fluor® 647 Conjugate

Cell Biology
University of Miami
RA3

Overall

Quality of Results

Ease-of-Optimization

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Company:

Life Technologies

Product Name:

Donkey anti-Mouse IgG (H+L) Secondary Antibody, Alexa Fluor® 647 conjugate

Catalog Number:

A-31571

This is a great product to do immunofluorescence. We study primary cells (airway epithelial cells) in our lab and can use several different donkey alexas with different fluorophore conjugations in a single staining without any issue. A dilution of 1:2000 for 30 to 60 minutes works great with most of them. The only issue is that usually when the product is running low or is old, like most conjugated antibodies, they tend to aggregate .

Experimental Design and Results Summary

Applications

Immunofluorescence

Sample

NHBE

Primary Incubation

Usually overnight at 4 degrees and dilution depends on primary Ab in 3% BSA

Blocking Agent

3% BSA

Secondary Incubation

1:2000 ; 30-60 minutes at room temperature in 3% BSA

Tertiary Incubation

NA

Detection

Confocal microscopy

Results Summary

Can use up to 3 different donkey AF (3 different species and 3 different fluorophores) without any issue in background or cross reactivity.

Additional Notes

None

Image Gallery

Summary

The Good

Easy to optimize, lasts forever and still works great even when old

The Bad

The only problem is, like most conjugated antibodies, they tend to aggregate when they are getting old or running low, however a quick spin will solve this issue.

The Bottom Line

Amazing product, strongly recommend

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