The AJ519 antibody labels the human TAC/IL2RA protein by immunofluorescence

“…The cells were fixed with paraformaldehyde and permeabilized with saponin. The IL2Ra fusion protein was detected simultaneously with an anti-IL2Ra antibody (AJ519, coupled to a rabbit Fc domain) [10] and a recombinant antibody recognizing the fused epitope tag (e.g., anti-HA fused to a mouse Fc domain; Fig. 1B).…”

Section: A Comparative Assessment Of Different Tagsmentioning

confidence: 99%

A quantitative comparison of antibodies against epitope tags for immunofluorescence detection

Marchetti,

Lima,

Hammel

et al. 2023

FEBS Open Bio

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Epitope tags recognized by specific antibodies have been widely used over the last few decades, notably to localize tagged proteins within cells by immunofluorescence. The diversity of tags and antibodies usually prevents a side‐by‐side comparison of the efficiency with which each antibody recognizes its cognate tag. We expressed chimeric proteins, each composed of an invariant domain (IL2Ra) associated with a specific epitope tag. Double immunofluorescence allowed us to quantify in parallel the reference signal generated by the anti‐IL2Ra antibody and the signal generated by the anti‐epitope tag antibody. Since all antibodies used in this study were recombinant antibodies fused to the same mouse Fc domain, the generated signals were directly comparable. Three groups of tags/antibodies were revealed: ‘good’ antibodies generated high signals even when used at a low concentration (50 ng·mL−1), ‘fair’ antibodies generated a high signal only at high concentrations (5000 ng·mL−1), and ‘mediocre’ antibodies generated positive but weak signals. Except for an anti‐myc antibody, similar results were obtained when cells were fixed in paraformaldehyde or methanol. These results provide a side‐by‐side quantitative evaluation of different tag/antibody pairs. This information will be useful to optimize the choice of epitope tags and to choose optimal antibodies.

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“…Supernatants (see Table 1 for individual yields) were collected after 4 days. The antibody AJ519, which recognizes the TAC antigen (human IL2RA, Uniprot P01589), was used as a negative control (Arsimoles et al, 2020). As positive control, a commercial anti-beta actin antibody (clone 2D4H5, Proteintech 66009-1-Ig), raised against human ACTB (Uniprot P60709), was used.…”

Section: Antibodiesmentioning

confidence: 99%

The AE765, AK423, AK692, AO233 and AO234 antibodies recognize human actin by immunofluorescence in HEK cells

Marchetti¹

2022

Antib. Rep.

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The AJ519 antibody labels the human TAC/IL2RA protein by immunofluorescence

Abstract: The recombinant antibody AJ519 detects by immunofluorescence the human TAC surface protein in paraformaldehyde-fixed cells.

Cited by 7 publications

References 4 publications

The AW288 and AX067 antibodies recognize human actin by immunofluorescence in HEK cells

The AW288 and AX067 antibodies recognize human actin by immunofluorescence in HEK cells

A quantitative comparison of antibodies against epitope tags for immunofluorescence detection

The AE765, AK423, AK692, AO233 and AO234 antibodies recognize human actin by immunofluorescence in HEK cells

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