AKT2 antibody | knockout validation | LSBio LS-C156232

This is a knockout-validated antibody summary, based on the publication "Immunohistological Examination of AKT Isoforms in the Brain: Cell-Type Specificity That May Underlie AKT's Role in Complex Brain Disorders and Neurological Disease", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

AKT2 antibody | knockout validation | LSBio LS-C156232 figure 1
Figure 1. Western blot analysis of AKT isoform levels in hippocampal lysates prepared from Akt mutant mice. Cre-mediated removal of floxed Akt2 under the early neural progenitor Nes promoter in an Akt3 KO background (n2F3K) led to near-complete removal of both AKT2 and AKT3 in the hippocampus (AKT2: t(5)=12.07, P<0.001; AKT3: t(5)=40.15, P<0.001). This genetic manipulation also led to a significant increase in the AKT1 levels in n2F3K mice when compared with WT controls (t(5)=6.471, P=0.001). Cre-mediated removal of floxed Akt1 under the excitatory neuron Cam2α promoter in an Akt3 KO background (c1F3K) led to a small reduction in AKT1 levels in the hippocampus (t(5)=4.310, P=0.008) with complete AKT3 loss (t(5)=16.99, P<0.001) but no change in AKT2 levels (t(5)=0.340, P>0.05). Nes-Cre-mediated removal of floxed Akt1 and Akt2 (n1F2F) resulted in near-total removal of AKT1 (t(6)=15.45, P<0.001) and AKT2 (t(6)=17.80, P<0.001) but no significant change in AKT3 expression (t(6)=0.297, P>0.05). N=3–4/genotype. From [1].
Antibody information

Mouse monoclonal IgG1

Company: LSBio

Antibody: AKT2

Catalog number: LS-C156232

Summary: Mouse monoclonal antibody to human AKT2 (aa214-451). Reacts with human and mouse. Suitable for western blot, immunofluorescence and immunohistochemistry.

Validation Method

Western blot

Sample

WT and AKT2-KO mouse hippocampus.

Blocking agent

0.2% I-Block (Tropix) in Tris-buffered saline with 0.1% Tween-20 (TBS-T) for 1 h at RT.

Primary incubation

1:1,000 dilution for up to 72 h at 4 °C.

Secondary incubation

1:5,000–1:20,000 dilution HRP-conjugated goat anti mouse secondary antibodies (Promega) in I-Block solution at RT for 1 h.

Detection

Enhanced chemiluminescence (GE Healthcare)

References
  1. Levenga J, Wong H, Milstead R, LaPlante L, Hoeffer C. Immunohistological Examination of AKT Isoforms in the Brain: Cell-Type Specificity That May Underlie AKT's Role in Complex Brain Disorders and Neurological Disease. Cereb Cortex Commun. 2021;2:tgab036 pubmed publisher