Dependable AP-Conjugated Anti-Mouse Antibody From Jackson Laboratories for IgG2c Isotype Specific Antibody Detection Using ELISA

December 29, 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 subclass 2c specific

Catalog Number:

115-055-208

Clone Number:

Polyclonal

I have used this secondary antibody to detect mouse antibodies of the IgG2c isotype, regarded to be generated from type 1 polarizing adjuvants such as Poly IC, from mouse serum, using an ELISA. The secondary specifically binds to antibodies of this isotype and is conjugated to an alkaline-phosphatase enzyme that generates color in the presence of the AP substrate. 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 IgG2c isotype specific antibodies, considered to be highly potent for anti-viral immunity, made in mice to Spike protein immunogen delivered to mice during subcutaneous immunizations. Mice were immunized with different adjuvants that polarize the antibody response to IgG1, IgG2b, IgG2c isotypes. 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 IgG2c isotype-specific 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, stable at 4 degrees C for months.

The Bad

Alkaline phosphatase enzyme reaction is highly sensitive to pH and temperature.

The Bottom Line

I would highly recommend this antibody for reproducible results when using the ELISA to detect antigen specific IgG2c isotype antibodies from mouse samples. Although slightly more expensive, I trust this antibody the most for its specificity and reliability.

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