unlabeled wave2 mouse c  (Santa Cruz Biotechnology)

Journal: Frontiers in cell and developmental biology

Article Title: Ena/VASP proteins at the crossroads of actin nucleation pathways in dendritic cell migration.

doi: 10.3389/fcell.2022.1008898

Figure Lengend Snippet: FIGURE 6 The VASP-EVH1 domain interacts with Wave2 and mDia1 in DCs. (A) Domain structure of Ena/VASP proteins and derived GST-fusion protein. EVH1, Ena/VASP homology 1 domain; EVH2, Ena/VASP homology 2 domain; PRR, Proline-rich region; GAB, G-actin binding site; FAB, F-actin binding site; GST: Glutathione S-transferase; FPPPP, Pro-rich motif recognized by EVH1 domain. (B–D) Recombinant GST-VASP-EVH1 and GST as control were purified from bacteria with GST-beads and incubated with detergent-extracted lysate of immature or mature DCs in the presence or absence of an EVH1-specific inhibitor. After thorough washing, proteins were eluted and analyzed by mass spectrometry (B,C) or by immunoblotting with the indicated antibodies (D). Volcano plots illustrate the inhibitor-sensitive hits obtained by mass spectrometry for iDCs (B) and mDCs (C). (D) Immunoblotting confirms that Abi1 and Wave2 (as representatives of the WRC) and mDia1 from iDCs and mDCs show specific binding to the EVH1 domain of VASP which is inhibited by the EVH1 domain-specific inhibitor.

Article Snippet: Antigen Species Clone name or catalog # Supplier IF WB FACS Comments α-Tubulin Mouse DM1A Cell signalling -- 1:500 -- unlabeled β-Actin Mouse Ac-15; A5441 Sigma-Aldrich -- 1:1,000 -- unlabeled CD11c Hamster N418; MCD11c05 Thermo Fisher Scientific -- -- 1:400 APC-labeled CD40 Mouse 3/23; #562846 BD Biosciences -- -- 1:400 Pacific Blue-labeled CD86 Rat GL1 eBioscience -- -- 1:100 APC-labeled EVL Rabbit TA343847 Origene -- 1:500 -- unlabeled GAPDH Mouse G8795 Sigma -- 1:1,000 -- unlabeled mDia1 Rabbit Ab129167; Ab96784 Abcam -- 1:1,000 -- unlabeled mDia1 Mouse Clone 51; 610,848 BD Biosciences 1:50 -- -- unlabeled Lamellipodin Rabbit -- Matthias Krause, Kings College London -- 1:20,000 -- unlabeled Mena Rabbit LS-C170270 LSBio -- 1:500 -- unlabeled p34/ARPC2 Rabbit 07-227-I Sigma-Aldrich 1:100 1:1,000 -- unlabeled Paxillin Rabbit Y113; ab32084 Abcam 1:400 -- -- unlabeled VASP Rabbit 9A2 Cell signalling -- 1:500 -- unlabeled Vinculin Mouse V9264 Sigma -- 1:1,000 -- unlabeled Vinculin Rabbit Ab73412 Abcam 1:400 -- -- unlabeled Wave2 Rabbit D2C8 Cell signalling -- 1:500–1:1,000 -- unlabeled Wave2 Mouse C-6; sc-373889 Santa Cruz Biotechnology 1:100 -- -- unlabeled Zyxin Rabbit Z4751 Sigma 1:400 1:500 -- unlabeled Frontiers in Cell and Developmental Biology frontiersin.org04 (Visweshwaran and Maritzen, 2019) with the following modifications: The migration chamber was coated with 10 μg/ml fibronectin for 1 h at 37°C using 150 μl of solution.

Techniques: Derivative Assay, Binding Assay, Recombinant, Control, Bacteria, Incubation, Mass Spectrometry, Western Blot

Journal: Frontiers in Oncology

Article Title: BTG1 Overexpression Might Promote Invasion and Metastasis of Colorectal Cancer via Decreasing Adhesion and Inducing Epithelial–Mesenchymal Transition

doi: 10.3389/fonc.2020.598192

Figure Lengend Snippet: Antibodies used for Western blot.

Article Snippet: WAVE2 (C-6) , Mouse , Santa Cruz Biotechnology.

Techniques: Western Blot