Rabbit anti pvav1 y160

Cells were centrifuged, supernatant was cleared, and the cells were re-suspended in lysis buffer as previously described [9 (link),33 (link),34 (link)]. Cells were placed on ice for 30 min and then centrifuged. The cell lysate was collected and moved to fresh tubes. Sample buffer was then added to the cell lysates and samples were heated for 5 min at 100 °C and then centrifuged at 10,000× g.
Western blots were performed as described [6 (link),9 (link),34 (link)].
The primary antibodies for WB, PTP, and Imaging were: Rabbit anti-pVAV1 (Y160) (Bio Source Cat# Bs44482, Invitrogen, Thermofisher); human anti-CD28 (BioLegend, London, United Kingdom Cat# 302934); Rabbit anti-SHP-1 (C-19) (Santa-Cruz Biotechnology, Heidelberg, Germany, Cat# sc-287); Mouse anti-GAPDH (0411) (Santa-Cruz Biotechnology, Heidelberg, Germany, Cat# sc-47724); and Mouse anti-WASp (D1) (Santa-Cruz Biotechnology, Heidelberg, Germany, Cat# sc-5300).
Secondary Antibodies were: Goat anti-Rabbit (Santa Cruz Biotechnology, Heidelberg, Germany, Cat# sc-2004) and Goat anti-Mouse (Jackson ImmunoResearch, West Grove, PA, USA, cat# 115-035-003).

Antibodies and their sources were as follows: mouse anti-PLCγ1 (Upstate), mouse anti-VAV1 (D7), rabbit anti-SHP-1 and mouse anti-GAPDH (Santa Cruz), rabbit anti-pSHP-1 (S591) (ECM Biosciences), rabbit anti-pPLCγ1Y783 (Bio Source), rabbit anti-pVAV-1Y160 (Bio Source), and mouse anti-PKC-θ (Santa Cruz). Secondary antibodies: goat anti-mouse (Sigma-Aldrich), goat anti-rabbit (Santa Cruz).