Key Reagents Table
| Reagent type | Designation | Source | Identifiers | Additional Information |
|---|---|---|---|---|
| B-cells from C57BL/6 mice | WT | Jackson Laboratories | 000664 | |
| B-cells from WASP−/− mice | WKO | Jackson Laboratories | 019458 | |
| B-cells from CD19Cre/+N-WASPflox/flox mice | cNKO | Lisa Westerberg laboratory | ||
| B-cells from N-WASPflox/flox mice | Flox control | Lisa Westerberg laboratory | ||
| B-cells from LifeAct-GFP mice | LifeAct-GFP | Roberto Weigert laboratory | ||
| B-cells from LifeAct-RFP mice | LifeAct-RFP | Klaus Ley laboratory | ||
| B-cells from GFP-NMIIA mice | GFP-NMIIA | Robert Adelstein laboratory | ||
| Reagent | 1,2-dioleoyl-sn-glycero-3-phosphocholine | Avanti Polar Lipids | 850375 P | 5 mM |
| Reagent | 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-cap-biotin | Avanti Polar Lipids | 870273 C | 50 μM |
| Reagent | Streptavidin | Jackson Immuno Research | 016-000-084 | 1 μg/ml |
| Antibody | F(ab’)2 fragment of goat IgG anti-mouse Ig(G+M) (polyclonal) | Jackson Immuno Research | 115-006-068 | - |
| Antibody | Cy3-Fab fragment of goat anti-mouse IgG+M (polyclonal) | Jackson Immuno Research | 115-167-020 | 2.5 μg per 1 × 106 cells |
| Chemical | 2-Mercaptoethylamine HCL | Thermo Fisher Scientific | 20408 | 50 mM |
| Reagent | EZ-Link™ Maleimide-PEG2-biotin | Thermo Fisher Scientific | A39261 | 20 mM per mM of protein |
| Commercial Kit | Alexa Fluor 546 antibody labeling kit | Thermo Fisher Scientific | A20183 | - |
| Ligand | Biotinylated holo-transferrin | Jackson Immuno Research | 015-060-050 | - |
| Antibody | Rat IgG2b anti-mouse CD90.2 (Thy1.2) (monoclonal) | Biolegend | 105351 | 1 μl per 2×106 cells |
| Inhibitor | CK-689 | Millipore Sigma | 182517–25MG | 50 μM |
| Inhibitor | CK-666 | Millipore Sigma | SML0006–5MG | 50 μM |
| Inhibitor | Wiskostatin | Millipore Sigma | W2270–5MG | 10 μM |
| Inhibitor | Blebbistatin | Cayman Chemicals | 13013 | 50 μM |
| Antibody | Rabbit IgG anti-mouse Arp2 antibody (polyclonal) | Abcam | ab47654 | 1:100 |
| Antibody | Rabbit IgG anti-mouse pWASP antibody (polyclonal) | Thermo Fisher Scientific | PA5–105572 | 1:100 |
| Antibody | Rabbit IgG anti-mouse pN-WASP antibody (polyclonal) | Thermo Fisher Scientific | PA5–105307 | 1:100 |
| Antibody | AF488-goat IgG anti-rabbit IgG antibody (polyclonal) | Thermo Fisher Scientific | A-11034 | 1:200 |
| Antibody | AF546-goat IgG anti-rabbit IgG antibody (polyclonal) | Thermo Fisher Scientific | A-11035 | 1:200 |
| Reagent | Acti-stain-488 Phalloidin | Cytoskeleton | PHDG1-A | 200 nM |
| Reagent | Acti-stain-555 Phalloidin | Cytoskeleton | PHDH1-A | 200 nM |
| Antibody | Rabbit IgG anti-mouse NMIIA antibody (polyclonal) | Abcam | ab75590 | 1:100 |
| Antibody | Rabbit IgG anti-mouse pCD79a (Y182) (monoclonal) | Cell Signaling Technology | 14732S | 1:100 |
| Antibody | Rabbit IgG anti-mouse pSyk (Y519/520) (monoclonal) | Cell Signaling Technology | 2710S | 1:100 |
| Antibody | Rabbit IgG anti-mouse pSHIP-1 (Y1020) (polyclonal) | Cell Signaling Technology | 3941S | 1:100 |
| Antibody | Rabbit IgG anti-mouse Syk (polyclonal) | Thermo Fisher Scientific | PA5–17812 | 1:100 |
| Antibody | Rabbit IgG anti-mouse SHIP-1 (polyclonal) | Thermo Fisher Scientific | PA5–115894 | 1:100 |