PEG hydrogel surfaces were modified with thiolated-proteins or peptides via thiol-ene reactions. To introduce thiol group to the protein, proteins were reacted with 2-iminothiolane HCl (Traut's Reagent, Thermo Scientific) following the manufacturer's protocol. NHS-ester fluorescent tags (Dylight488 or Dylight594, Invitrogen) were also reacted with proteins/peptides for pattern visualization. Acrylate-acrylate and thiol-ene PEG hydrogels were photopolymerized, using the method described in the previous section. Next, reduced thiolated proteins or peptides were evenly pipetted across the surface of the hydrogel. For patterning, a photomask was placed directly onto the hydrogel surface. Hydrogels underwent a second round of UV exposure, for 30 s, unless otherwise stated. Hydrogels were placed into well plates for imaging or cell seeding. All hydrogels were washed in phosphate buffer saline (PBS) overnight prior to imaging or cell seeding.
2 iminothiolane hcl
Manufactured by Thermo Fisher Scientific
2-iminothiolane HCl is a reagent used for the introduction of sulfhydryl groups into biomolecules. It can be used for the covalent attachment of molecules to proteins and other macromolecules.
Automatically generated - may contain errors
Lab products found in correlation
Dylight 594, by Thermo Fisher Scientific (2 mentions)
Dylight 488, by Thermo Fisher Scientific (1 mentions)
Poloxamer 188, by Thermo Fisher Scientific (1 mentions)
Dapi 4 6 diamidino 2 phenylindole, by Thermo Fisher Scientific (1 mentions)
Rnase free water, by Thermo Fisher Scientific (1 mentions)
Fetal bovine albumin, by Thermo Fisher Scientific (1 mentions)
N p maleimidophenyl isocyanate pmpi, by Thermo Fisher Scientific (1 mentions)
3 protocols using 2 iminothiolane hcl
1
Patterned Protein-Functionalized PEG Hydrogels
2
Protein-Functionalized PEG Hydrogel Patterning
3
Neurotensin Receptor 1 Targeting Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Human IgG was purchased from Equitech Bio (Kerrville, TX, USA). Poloxamer-188, RNase-free water, 4,6-diamidino-2-phenylindole (DAPI), and fetal bovine albumin (FBS) were obtained from Fisher Scientific. Antineutotensin receptor 1-monoclonal antiboby (anti-NTSR1-mAb was purchased from Santa Cruz Biotechnologies. siRNA against mutated KRAS G12S was designed andpurchased from GE Dharmacon. siG12S sense and antisense sequences are GUUGGAGCUAGUGGCGUAGdTdT and CUACGCCACUAGCUCCAACdTdT respectively. siGLO-Green (6-FAM-labeled) was obtained from GE Dharmacon. N-[p-maleimidophenyl] isocyanate (PMPI), and 2-iminothiolane-HCl were obtained form Thermo Scientific. Neurotensin was purchased from Abcam (Cambridge, MA, USA)
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