Description
General description
The Atlas MicroMap Kit is a highly precise proximity labeling platform for elucidation of protein-protein interactions on cell membranes. Using an antibody specific to a protein target, the Atlas MicroMap kit crosslinks proteins within a 4 nm radius to identify only those that are within the proximity of the target of interest. The kit contains necessary reagents for performing photoinduced proximity labeling using the Atlas system. The procedures are adopted from those published by the MacMillan Lab in Science . Two sizes of kit are available providing enough antibody conjugate for either 10 reactions (AtlasKit-A) or 20 reactions (AtlasKit-B).
The kit provides a step-by-step procedure for conjugating your antibody to a photocatalyst that localizes light-mediated crosslinking to the antibody′s target and surrounding proteins. First, an azide-containing linker is appended to your antibody that ″clicks″ on an iridium photocatalyst. The resulting antibody-photocatalyst conjugate is added to live cells along with a biotinylated diazirine reagent. Analogous to a satelite, the photocatalyst localizes carbene generation from the diazirine and thus covalently modifies surrounding proteins. The biotin tag allows for enrichment of photo-labeled proteins. As shown in Geri et al., the extent of this photo-induced labeling is within a 4 nm radius of the photocatalyst, providing advanced precision for mapping protein interaction networks.
Read more about micromapping in our Technology Spotlight
Protocol available here
Included:
Conjugation reagents and buffers, photocatalyst, photo-reactive diazirine-biotin, fluorescent standard for validation
Not included:
Antibodies (primary, secondary, isotype)
Light-irradiating photoreactor (suggestions below)
Protein concentration assay (BCA assay recommended)
Plate reader
Spin desalting columns (7K MWCO, 2 mL recommended)
Photoreactors commercially available:
Penn PhD Photoreactor M2 (Z744035)
Penn PhD Photoreactor M2 365nm Light Source (Z744031)
PennPhd Lightsource to 450nm Light Source (Z744033)
Application
Proximity protein labeling, protein-protein interactions, photoaffinity labeling, proteomics
Biochem/physiol Actions
Kit of components packaged in individual vials for enabling photoinduced proximity labeling. Kit includes reagents for antibody conjugation, click chemistry, and photo-labeling of protein. Chemistry is compatible with customer-specific antibody (not supplied as part of kit) for proteins of interest. Kits are available for either 10 reactions (AtlasKit-A) or 20 reactions (AtlasKit-B). A reaction is defined as a single antibody conjugation preparation.
