This is a knockout-validated antibody summary, based on the publication "The influence of AKT isoforms on radiation sensitivity and DNA repair in colon cancer cell lines", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

Figure 1. Western blots of AKT1 in the colorectal cancer cell lines DLD-1 and HCT116 in the parental WT, AKT1-KO, AKT1-KO and AKT1/2 double KO cells. Beta-actin is used as loading control. From [1].

Mouse monoclonal IgG1 (kappa light chain)

Company: Santa Cruz Biotechnology

Antibody: AKT1

Catalog number: sc-55523

Summary: Mouse monoclonal IgG1 (kappa light chain) raised against amino acids 345-480 of Akt1 of human origin. Recommended for detection of Akt1 of mouse, rat and human origin by western blot, immunoprecipitation, immunofluorescence and ELISA

Western blot

WT, AKT1-KO and AKT1/2 double KO DLD-1 and HCT116 cells, in lysis buffer containing 1 % Tween-20, 20 mM Tris (pH 8.0), 137 mM NaCl, 10 % glycerol, 2 mM EDTA, 1 mM activated sodium orthovanadate (Sigma), and protease inhibitor cocktail (P8340, Sigma)

1 h in 5 % BSA, PBS

overnight at 4 °C

horseradish peroxidase-labeled secondary antibody (626520) (Invitrogen) for 1 h at room temperature

CCD camera (SuperCCD HR, Fujifilm, Japan) after treatment with electrochemiluminescent solution (Immobilon) for 5 min.

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.