Well Working Secondary Antibody From Thermo Fisher for Immunofluorescence Detection (405 nm laser)

February 06, 2024

IKP/RBCT
Robert Bosch Gesellschaft für medizinische Forschung mbH
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

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Company:

ThermoFisher Scientific

Product Name:

Goat anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor™ Plus 405

Catalog Number:

A-48255

Clone Number:

This secondary anti-mouse antibody from Thermo Fisher Scientific (Invitrogen) is a good choice for the detection of primary mouse antibodies, for example, for immunofluorescence in the 405 nm range (UV laser). The antibody can be easily combined with multiple additional stains in the green, red, or far-red range.

Experimental Design and Results Summary

Applications

Immunofluorescence

Sample

Formaldehyde-fixed and permeabilised (human) cells incubated with a primary mouse antibody

Primary Incubation

1:100, 2h, RT

Blocking Agent

2 % BSA + 1 % FCS in PBS

Secondary Incubation

1:100, 1h, RT

Tertiary Incubation

None

Detection

Confocal laser scanning microscopy (405 nm laser)

Results Summary

Detection with the Alexa Fluor Plus 405 give a strong signal when excited with a UV-laser and can be easily separated from green (488 nm), red (594nm) or far-red (638nm) dyes. Signal intensity mainly depends on quality of the primary antibody used.

DOI or PMID #

-

Additional Notes

Store the antibody at 4°C, stable for several weeks even months, test with a positive control (established primary mouse antibody before use).

Image Gallery

Summary

The Good

Bright and stable signal.

The Bad

Pricey, sometimes optimization needed (antibody-dependent).

The Bottom Line

This Alexa Fluor Plus 405 antibody for detection of primary mouse antibodies gives a bright and stable signal in combination with a 405 UV laser. Optimization for different primary antibodies possibly needed, however, works fine in our hands for IF.

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