Anti rabbit igg h l irdye800

Anti-Total pY (clone 4G10), 1:1000, Millipore # 05-321; anti-Btk (D6T2C), 1:1000, Cell Signaling # 56044; anti-Btk, 1:1000, Thermo Scientific # PA5-27392; anti-Btk (pY551)/Itk (pY511) Clone 24a, 1:1000, BD Biosciences # 558034; anti-Btk (pY223), 1:1000, Cell Signaling #5082; anti-p44/42 MAPK (Erk1/2), 1:1000, Cell Signaling #9102; anti-phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (E10), 1:1000, Cell Signaling #9106; anti-PLCƴ2, dilution 1:1000, Cell Signaling, #3872; anti-PLCƴ2 (Tyr1217), 1:1000, Cell Signaling #3871; anti-Flag, 1:5000, Sigma # F3165; anti-Penta-his, 1:5000, Qiagen #34660; anti-PLCƴ2 (Tyr753) [EPR5914-3], 1:1000, Abcam #ab133455; anti-Tubulin, 1:2000, Sigma #T9026; anti-Myc-tag Myc.A7 DyLight800, 1:10,000, Thermo Scientific #MA1-21316-D800; anti-mouse IgG IRDye 800CW, 1:10,000, LiCor #926-32210; anti-Rabbit IgG (H+L) IRDye800, 1:10,000, Rockland #611-732-127; anti-Mouse IgG (H+L) Peroxidase AffiniPure, 1:10,000, Jackson ImmunoResearch #115-035-003; anti-Rabbit IgG (H+L) Peroxidase AffiniPure, 1:10,000, Jackson ImmunoResearch #111-035-003. The full information on primary and secondary antibodies that were used for the reported experiments is available in the Reporting Summary.

For detection of proteins in western blot the following primary antibodies were used: the mouse monoclonal anti-fibronectin C6F10 (1:200 dilution), the rat monoclonal anti-nidogen ELM1 (1:500 dilution) (sc-73611, sc-33706; Santa Cruz, Heidelberg, Germany), the rabbit polyclonal anti-collagen type IV (1:200 dilution) (AB756P; Millipore, Darmstadt, Germany) and the rabbit polyclonal anti-laminin (1:500 dilution) (L9393; Sigma-Aldrich, St.Louis, USA). The following secondary antibodies were used: anti-mouse IgG, HRP-linked (1:2000 dilution) (7076; Cell Signaling Technologies, Cambridge, UK), anti-rabbit IgG (H+L), HRP-linked (1:2000 dilution) (111–035–144; Dianova, Hamburg, Germany), anti-rat IgG (H+L), HRP-linked (1:2000 dilution) (6180–05; SouthernBiotech, Birmingham, USA) and anti-rabbit IgG (H+L), IRDye 800 (1:5000 dilution) (611–132–122; Rockland, Gilbertsville, USA). The ECM gels were thawed on ice and heated in Laemmli buffer at 95°C for 5 min. Equal amounts of protein were loaded on polyacrylamide gels. The proteins were separated by SDS-PAGE and transferred to nitrocellulose membranes using tank blotting. For the detection of protein levels, the ECL detection system (Amersham Pharmacia Biotech, Uppsala, Sweden) or Odyssey Infrared system version 2.1 (LI-COR Biosciences, Lincoln, USA) was used.