This is a knockout-validated antibody summary, based on the publication "Postnatal developmental expression of regulator of G protein signaling 14 (RGS14) in the mouse brain", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Mouse monoclonal IgG2a

Company: Neuromab

Antibody: RGS14

Catalog number: 75-170

Summary: Mouse monoclonal IgG2a against a fusion protein corresponding to full-length rat RGS14. Reacts with mouse and rat. Suitable for western blot, immunocytochemistry and immunohistochemistry.

Western blot | Immunohistochemistry

Brains from wild-type and RGS14-knockout mice. WB: brains were homogenized in ice-cold homogenization buffer containing 50 mM Tris, 150 mM NaCl, 1 mM EDTA, 2 mM DTT, 5 mM MgCl2, phosphatase inhibitors (1:1,000, Sigma Aldrich), and one mini protease inhibitor cocktail tablet (Roche Applied Science, Basel, Switzerland), diluted with dH2O to 10 mL, pH 7.4. IHC: Brains were fixed in 4% PFA, embedded in paraffin and coronally sliced in 10 µm thick sections.

WB: blocking buffer (5% nonfat milk (w/v), 0.1% Tween-20, and 0.02% sodium azide, diluted in 20 mM Tris buffered saline, pH 7.6) for 1 hour at room temperature.

IHC: 1:250 dilution goat anti-mouse IgG AffiniPure fab fragment (Jackson Immunoresearch) in blocking buffer (Vectastain standard ABC kit, Vector Laboratories). for 1 hour and 45 minutes at 4°C.

WB: 1:1,000 dilution in blocking buffer overnight at 4°C.

IHC: 1:500 dilution in Tau Secret Formula for 24 hours at 4°C.

WB: 1:5,000 dilution HRP-conjugated anti-mouse secondary antibody in TBST.

IHC: 1:200 dilution horse anti-mouse biotinylated secondary antibody in Tris Brij containing 2% horse serum for 30 minutes at 37°C.

WB: enhanced chemiluminescence and exposing membranes to films.

IHC: avidin-biotin peroxidase complex (ABC) solution (Vectastain standard ABC kit, Vector Laboratories) for 80 minutes at 37°C and 0.096% 3,3’-diaminobenzidine tetrahydrochloride (DAB, Vector Laboratories) for 5 minutes. Images were obtained with an Olympus BX51 light microscope, using DP Controller software on a DP70 camera (Olympus).

Immunoblotting of RGS14 in wild-type (WT) and RGS14 knockout (KO) adult mouse brain homogenates. Beta-actin was used as loading control. Immunohistochemistry of RGS14 in hippocampus at different developmental stages and wild-type (WT) and RGS14 knockout (KO) adult mouse brain. Scale bar = 1.0 mm. Please see Figure 2b,c in the article [1]