Key resources table.
| Reagent type (species) or resource |
Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| Gene (human) | CaMKII-α | Uniprot_ID: Q9UQM7 | ||
| Gene (human) | CaMKII-β | Uniprot_ID: Q13554 | ||
| Gene (human) | CaMKII-β’E | Uniprot_ID: Q13554-3 | ||
| Gene (human) | CaMKII-γ | Uniprot_ID: Q13555 | ||
| Gene (human) | CaMKII-δ | Uniprot_ID: Q13557 | ||
| Recombinant DNA reagent | pET21a-BirA | Addgene | plasmid # 20857 | for biotinylation of the AVI tag in HEK 293T cells |
| Recombinant DNA reagent | EYFP-CaM | Addgene | plasmid # 47603 | coexpressed for in cell activation of CaMKII |
| Recombinant DNA reagent | pEGFP-C1 (plasmid) | Clontech, Mountain View, CA | vector backbone for inserting the CaMKII genes | |
| Recombinant DNA reagent | pSNAPf (plasmid) | New England Biolabs, MA | N9183S | vector backbone |
| Cell line (human) | HEK 293T | UC Berkeley cell culture facility | authenticated using STR profiling and tested negative for mycoplasma | |
| Antibody | anti-CaMKII (phospho T286); mouse monoclonal | Abcam | ab171095 | 1:500 |
| Antibody | anti-CaMKII (phospho T306); rabbit polyclonal | PhosphoSolutions | p1005-306 | 1:500 |
| Peptide, recombinant protein | Poly-L-lysine PEG (PLL:PEG) | SuSoS, Dübendorf, Switzerland | PLL(20)-g[3.5]- PEG(2) | preparation of flow chambers |
| Peptide, recombinant protein |
streptavidin | Sigma-Aldrich | S0677 | functionalize the glass substrates for capturing biotinylated CaMKII |
| Peptide, recombinant protein | calmodulin | Sigma-Aldrich | C4874 | activation of CaMKII |
| Peptide, recombinant protein |
λ-phosphatase | New England Biolabs, MA | P0753L | phosphatase |
| Peptide, recombinant protein | PP1α | EMD Millipore, Burlington, MA |
14–595 | phosphatase |
| Chemical compound, drug | PEG-Biotin | SuSoS, Dübendorf, Switzerland | PLL(20)-g[3.5]- PEG(2)/PEG(3.4)- biotin(50%) | preparation of flow chambers |
| Chemical compound, drug | 1% protease inhibitor cocktail | Sigma | P8340 | protease inhibitor cocktail for lysis buffer |
| Chemical compound, drug | 0.5% phosphatase inhibitor cocktail 2 and 3 | Sigma | P0044 and P5726 | phosphatase inhibitor cocktails for lysis buffer |
| Chemical compound, drug | staurosporine | Abcam | ab120056 | kinase inhibitor |
| Chemical compound, drug | cyclosporin A | Sigma-Aldrich | 30024 | phosphatase inhibitor |
| Chemical compound, drug | okadaic acid | Abcam | ab141831 | phosphatase inhibitor |
| Software, algorithm | FIJI (ImageJ) | open access software, see https://imagej.net/Fiji/Downloads | image processing | |
| Software, algorithm | in-house Matlab codes | open access, see Source code 1 | image processing | |
| Other | sticky-Slide VI 0.4 | Ibidi | 80608 | flow chambers |
| Other | glass coverslips | Ibidi | 10812 | functionalized substrates |