This is a knockout-validated antibody summary, based on the publication "Repeated inhalation of sevoflurane inhibits the information transmission of Purkinje cells and delays motor development via the GABAA receptor ε subunit in neonatal mice", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

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Figure 1. Western blot showing the expression levels of the GABAA-Rε subunit in the KO and WT mice. From [1].

Mouse monoclonal IgG1 (kappa light chain)

Company: Santa Cruz

Antibody: GABAA Rε

Catalog number: sc-271668

Summary: Mouse monoclonal IgG1 (kappa light chain) raised against amino acids 1-110 mapping at the N-terminus of human GABAA Rε. Recommended for detection of GABAA Rε of mouse, rat and human by western blot, immunoprecipitation, immunofluorescence and ELISA.

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Figure 2. Immunohistochemistry of cerebellar sections from GABAA-Rε subunit KO and WT mice. GABAA-R ε subunit (red), pan neuronal marker (green) and DAPI were stained and merged. Scale bar, 50 µm. From [1].

Western blot | Immunohistochemistry

Cerebellum from WT and GABAA-Rε KO mice. WB: Total proteins were extracted using radioimmunoprecipitation assay lysis solution (Beyotime Institute of Biotechnology). IHC: cerebellum tissue was fixed in 4% paraformaldehyde (Beyotime Institute of Biotechnology), followed by 30% sucrose (Sangon Biotech Co., Ltd., Shanghai, China; diluted in PBS) and embedded in OTC (Sangon Biotech Co., Ltd.) for cryosectioning.

WB: 5% bovine serum albumin (Beyotime Institute of Biotechnology) diluted with PBS-Tween-20 (PBST) at room temperature for 1 h.

IHC: 10% goat serum (OriGene Technologies, Inc., Beijing, China) diluted in 0.3% Triton X-100/0.1 M PBS at room temperature for 30 min.

WB/IHC: 1:300 dilution at 4˚C overnight.

WB: 1:3,000 dilution mouse IgG BP conjugated to CruzFluor™ 488 (sc‑516176; Santa Cruz Biotechnology, Inc.) at 37˚C for 1 h.

IHC: 1:500 dilution goat anti-mouse IgG fluorescein isothiocyanate (F‑2761; Invitrogen; Thermo Fisher Scientific, Inc., Waltham, MA, USA) or 1:500 dilution goat anti-mouse IgG phycoerythrin (P‑852; Invitrogen; Thermo Fisher Scientific, Inc.) for 2 h at room temperature.

WB: enhanced chemiluminescence solution (Beyotime Institute of Biotechnology) and scanned by Chemidoc XRS+ Imager (Bio-Rad Laboratories, Inc., Hercules, CA, USA).

IHC: All the samples were observed by fluorescence microscope (magnification, x400) (Zeiss AG, Oberkochen, Germany).