To investigate the coupling of lysine containing proteins into the
gelPEG hydrogel network, laminin 521 (Biolamina, Sweden) served as a model
protein. laminin 521 (LN) was added at a concentration of 10 μg mL−1to the reaction mixture of 3% w/v 4:1 gelPEG in TBS (n = 5
technical replicates). Hydrogels without LN served as a control
(n = 5 technical replicates). Hydrogel discs (≈1 × 5 mm) of
20 μL volume were incubated for 24 h at 37 °C in TBS. All hydrogels were washed
and fixed in 4% formalin and stained with a primary anti-LN α5
antibody (1:260, clone 4C7, MAB1924, Merck), followed by a goat-antimouse
antibody Alexa Fluor 546 (4 μg mL−1, A-11 003, Thermofisher). Imaging
of control and LN-laden hydrogels occurred with a fluorescence microscope (B×51,
Olympus).
gelPEG hydrogel network, laminin 521 (Biolamina, Sweden) served as a model
protein. laminin 521 (LN) was added at a concentration of 10 μg mL−1to the reaction mixture of 3% w/v 4:1 gelPEG in TBS (n = 5
technical replicates). Hydrogels without LN served as a control
(n = 5 technical replicates). Hydrogel discs (≈1 × 5 mm) of
20 μL volume were incubated for 24 h at 37 °C in TBS. All hydrogels were washed
and fixed in 4% formalin and stained with a primary anti-LN α5
antibody (1:260, clone 4C7, MAB1924, Merck), followed by a goat-antimouse
antibody Alexa Fluor 546 (4 μg mL−1, A-11 003, Thermofisher). Imaging
of control and LN-laden hydrogels occurred with a fluorescence microscope (B×51,
Olympus).