Reliable AP-conjugated Anti-Mouse IgG Antibody for ELISA From Jackson ImmunoResearch Laboratories

December 26, 2023

Department of Immunology and Microbiology
Scripps Research
Graduate Student

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Company:

Jackson Labs

Product Name:

Alkaline Phosphatase AffiniPure Goat Anti-Mouse IgG, Fcγ fragment specific

Catalog Number:

115-055-071

Clone Number:

Polyclonal

I have used this secondary antibody to detect mouse IgG antibodies from mouse serum in an ELISA setting. The secondary is conjugated to an alkaline-phosphatase enzyme that generates color in the presence of the AP substrate. I've used this antibody for more than 2 years and got reliable, reproducible results every time. I would highly recommend this secondary antibody.

Experimental Design and Results Summary

Applications

ELISA

Sample

Mouse serum, BALF

Primary Incubation

Anti-his antibody coating

Blocking Agent

3% BSA in PBS

Secondary Incubation

Diluted serum (in 1% BSA PBS-t)

Tertiary Incubation

This secondary antibody (in 1% BSA PBS-t)

Detection

Plate reader for 405 nm

Results Summary

I used this product to identify antibodies made in mice to Spike protein immunogen delivered to mice during subcutaneous immunizations. Serum or BALF was collected from immunized mice at various time points. ELISA half-well high binding plates were coated with 50 ul of Rabbit anti-his antibody overnight. The plates were then blocked and coated with his-tagged Spike protein. Plates were then washed and serum/BALF sample was added. Finally, the secondary antibody was added and used to quantify the level of anti-Spike antibodies of various isotypes present in the mouse sample.

DOI or PMID #

N/A

Additional Notes

N/A

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Summary

The Good

Reliable, cheap, and stable at 4 degrees C for months.

The Bad

Reaction is very sensitive to temperature and pH of the enzyme substrate.

The Bottom Line

I would highly recommend this antibody for reproducible results when using the ELISA to detect antigen specific antibody titers from mouse samples. This antibody allows for high sensitivity of detection.

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