Great Antibody To Stain mTOR For Confocal Microscopy

Oncology
Johns Hopkins School of Medicine
Graduate Student

Overall

Quality of Results

Ease-of-Optimization

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Company:

Cell Signaling Technology

Product Name:

mTOR (7C10) Rabbit mAb

Catalog Number:

2983

mTOR sub cellular localization is very critical for it's activation. mTORC1 activation takes place at the lysosomal surface of cells. During nutrient replete conditions, mTOR is readily observed at the lysosomal surface; however, during depleting conditions, mTOR is diffused throughout the cell. We were interested in observing this phenomenon in T cells during activation. This antibody is strong monoclonal antibody used by the mTOR field with immunofluorescence studies.

Experimental Design and Results Summary

Applications

Immunofluorescence

Sample

Murine CD8 T cells

Primary Incubation

1:400. Overnight at 4 deg or 2hr at room temperature

Blocking Agent

5% Goat Serum in PBS (.1% Tween)

Secondary Incubation

Goat anti-Rabbit AF 647 or 488

Tertiary Incubation

None

Detection

Zeiss LSM780 confocal microscope

Results Summary

We were able to readily identify mTOR localization at the lysosomes in both nutrient replete and deplete conditions. We also observed more localization upon T cell activation.

DOI or PMID #

27064374

Additional Notes

N/A

Related Categories

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Summary

The Good

Great monoclonal antibody against mTOR at great cost

The Bad

None really

The Bottom Line

Highly recommend this antibody for IF experiments to identify mTOR at the subcellular level.

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