This is a knockout-validated antibody summary, based on the publication "Deconstructing Ras Signaling in the Thymus", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Company: Santa Cruz

Catalog number: sc-256

Summary: rabbit polyclonal, against the C-terminus of Sos 1 of human origin.

Dilution: 1:500

Figure 1. Western blot for Sos1 protein in purified CD4+CD8+ thymocytes from wild-type and SOS1-/- mice from figure 3 [1].

Catalog number: sc-15358

Summary: rabbit polyclonal, epitope corresponding to amino acids 1091-1170 mapping near the C-terminus of Sos 2 of human origin.

Dilution: 1:200

Figure 2. Western blot for Sos2 protein in purified CD4+CD8+ thymocytes from wild-type and SOS1-/- mice from figure 3 [1].

Catalog number: sc-8430

Summary: Mouse monoclonal IgG1 raised against full length RasGRP of rat origin. Recommended for detection of RasGRP of mouse and rat origin by western blot, immunoprecipitation, immunofluorescence and ELISA.

Dilution: 1:500

Figure 3. Western blot for RasGRP1 protein in purified CD4+CD8+ thymocytes from wild-type and SOS1-/- mice from figure 3 [1].

Note: a nonspecific minor band with a higher molecular weight.

WB, western blot.

purified CD4+CD8+ thymocytes from wild-type or knockout mice, solubilized in 2× sodium dodecyl sulfate (SDS) sample buffer containing 100 mM dithiothreitol.

4°C overnight.

horseradish peroxidase-conjugated antibodies (1:20,000; Millipore) at room temperature for 1 h.

Enhanced chemiluminescence was used to visualize protein products (Super Signal West Pico and Super Signal West Femto from Pierce).