This is a knockout-validated antibody summary, based on the publication "RasGRP3 limits Toll-like receptor-triggered inflammatory response in macrophages by activating Rap1 small GTPase", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Figure 1. RAW264.7 cells cotransfected with the guide RNA-encoding plasmid and Cas9-encoding plasmid were sorted and selected for single clones. The depletion of RasGRP3 was examined by western blotting. β-actin was used as loading control. From [1].

Rabbit monoclonal IgG

Company: Cell Signaling

Antibody: RasGRP3

Catalog number: 3334

Summary: Rabbit monoclonal IgG against a synthetic peptide corresponding to residues near the carboxy terminus of human RasGRP3. Reacts with human and mouse by western blot. It is predicted to react based on 100% sequence homology with monkey.

Western blot

Wild type and RasGRP3-KO RAW264.7 cells. Total cell lysates were prepared by using cell lysis buffer (Cell Signaling Technology) containing phosphatase inhibitor cocktail (Sigma).

100 ng ml−1 diluted in a buffer containing 50 mM Tris.HCl (pH 7.4), 150 mM NaCl and 0.05% Tween 20.

100 ng ml−1 diluted in a buffer containing 50 mM Tris.HCl (pH 7.4), 150 mM NaCl and 0.05% Tween 20.

Supersignal West Femto Maximum Sensitivity substrate (Pierce) and Syngene Bio Imaging Systems (Frederick, MD).

If the antibody described in this summary is a polyclonal antibody, since polyclonal antibodies are of limited quantity, please inquire the supplier whether any current polyclonal antibody with the same catalog number is exactly the same as the one described in this summary. Sometimes, different bleeds or different animals are used, usually with a different lot number. In such cases, the result in this summary may not apply to the new antibody with the same catalog number.