Micropatterned agarose gels for stamping arrays of proteins and gradients of proteins.
We describe a method for repetitive and rapid formation of planar microarrays and gradients of proteins using patterned agarose stamps. It demonstrates: (i) micropatterning of agarose gels with feature sizes as small as 2 microm; (ii) inking of posts (diameter 50-1000 microm) on patterned agarose stamps with one or multiple (here, eight) proteins and repetitive stamping of patterns (>100 times in the case of one protein) and arrays (20 times in the case of eight proteins) without the need for intermediate re-inking; (iii) transferring spots of proteins with good homogeneity in surface coverage to glass slides; (iv) applying this technique to surface-based immunoassays; (v) stamping that requires only sub-nanomolar amounts of protein (typically approximately 3 microg in approximately 0.6 microL of solution); (vi) stamping without the need for drying of the proteins, as opposed to stamping with stamps made of poly(dimethylsiloxane); and (vii) patterning gradients of proteins by allowing two proteins to diffuse toward each other in an agarose stamp, followed by printing the protein gradients onto a surface.
Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA 02138, USA.
Michael
M
Mayer
Jerry
J
Yang
Irina
I
Gitlin
David H
DH
Gracias
George M
GM
Whitesides
D005782:Gels / Q000737:chemistry*; D007118:Immunoassay / Q000295:instrumentation / Q000379:methods; D007281:Ink; D008856:Microscopy, Fluorescence; D040081:Protein Array Analysis / Q000295:instrumentation / Q000379:methods; D011506:Proteins / Q000032:analysis*; D012685:Sepharose / Q000737:chemistry*; D013499:Surface Properties
D005782:Gels; D011506:Proteins; D012685:Sepharose