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2 protocols using anti c cbl 2747

1

Molecular Mechanisms of DSG1 Regulation

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DSG1 and HA-tagged S100A4, CYP11A1, ACSS1, LY6G6D, NR5A1, AIF1L, and FAM167A were cloned into the MigR1 vector. pClneo-Myc-Erbin was obtained from Addgene. pFLAG-NIK-DN was described previously [30 (link)]. Anti-FAM167A (sc-393999), anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (sc-166574), anti-p100/p52 (sc-7386 X), anti-DSG1 (sc-59904), and anti-ubiquitin (sc-8017) antibodies were purchased from Santa Cruz Biotechnology; anti-NIK (4994), anti-p100/p52 (4882), anti-Myc (2276), anti-TRAF3 (4729), anti-CHIP (2080), and anti-c-cbl (2747) antibodies were purchased from Cell Signaling Technology; the anti-DSG1 antibody (32–6000) was purchased from Invitrogen; the anti-Erbin antibody (NBP2–13968) was purchased from Novus Biologicals; and the control IgG (H9658) was purchased from Sigma-Aldrich. Imatinib (sc-202180). DSG1-specific short hairpin RNA (shRNA) lentiviral particles (sc-35224-V) and control shRNA lentiviral particles (sc-108080) were obtained from Santa Cruz Biotechnology. Nilotinib (1750) was obtained from BioVision, recombinant FAM167A (MBS1363522) was obtained from MyBioSource, SN50 (481480) and MG132 (474790) were obtained from Sigma-Aldrich, and the Pierce Silver Stain Kit (24612) was obtained from Thermo Fisher Scientific.
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2

Immune Cell Activation Pathway Profiling

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Anti-CD3 clone 2C-11, anti-CD28 clone PV1, and LFA-1 blocking anti-CD11a clone M17/4 were obtained from BioXCell. Anti-pTyr clone PY-20 and anti-p85 (ABS234) was from Upsate (Millipore). Anti-HEF1 (CasL) clone 2G9 and anti-Pyk2 clone YE353 were obtained from Abcam. Anti-Rac1 (610650) and anti-AKT (559028) were from BD. Anti-pAKT (4051), anti-pERK (9101), and anti-c-Cbl (2747) were from Cell Signaling. Anti-Cdc42 (SC-87), anti-CrkL (SC-319), and anti-Cbl-b (SC-8006) were from Santa Cruz. Secondary antibodies conjugated to appropriate fluorophores were obtained from Molecular Probes and Jackson Immunoresearch. AlexaFluor-conjugated phalloidin was purchased from Molecular Probes. Recombinant mouse ICAM-1-Fc, SDF-1α, and P-selectin were purchased from R&D Systems. The ROCK inhibitor Y-27632, the pan PI3K inhibitor LY-294002, and the PI3Kδ inhibitor IC87114 were obtained from CalBiochem. The myosin inhibitor (s)-nitro-blebbistatin was purchased from Cayman Chemicals. The actin destabilizing drug latrunculin B, the Src inhibitor PP2, the Abl kinase inhibitor STI-571, and the PI3Kγ inhibitor CZC24832 were purchased from Sigma.
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