Goat Anti-Rat AF647 Is a Great Secondary Antibody for Immunofluorescence

November 09, 2015

Pathology
Stanford
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

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Company:

Thermo Fisher Scientific, Invitrogen

Product Name:

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

Catalog Number:

A-21247

Info: View Product View Product Specs

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When I stain tissue sections with a rat-derived primary antibody, I utilize this product to visualize the rat primary antibody stain in the AF647 channel.

Experimental Design and Results Summary

Applications

Immunofluorescence

Sample

Pre-fixed frozen mouse small intestine tissue section

Primary Incubation

1:100 in 1% BSA in PBS overnight at 4 degrees

Blocking Agent

5% goat serum in 1% BSA in PBS

Secondary Incubation

Goat anti-rat AlexaFluor647 was used at 1:200 in PBS and was incubated on the section for 1 hour at room temperature in the dark

Tertiary Incubation

NA

Detection

Zeiss LSM 700 confocal

Results Summary

The anti-rat stain (shown in white) shows good staining of the rat anti-mouse Foxp3 primary antibody. Here, CX3CR1-GFP is shown in green, anti-Foxp3 is shown in white (the AF647 stain), and anti-CD3 is shown in red (AF594).

Additional Notes

The AF647 fluorophore is good for triple color staining, but strong staining may bleed into the AF594 channel depending on filters.

Image Gallery

Summary

The Good

Stains the primary rat antibody very well

The Bad

May bleed into other channels

The Bottom Line

Overall, a great secondary antibody to use for triple -color stainings, or just for visualization of a rat primary antibody in the AF647 channel.

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