This is a knockout-validated antibody summary, based on the publication "Active and dynamic mitochondrial S-depalmitoylation revealed by targeted fluorescent probes", as cited below [1]. Labome curates formal publications to compile a list of antibodies with unambiguous specificity within Validated Antibody Database (VAD).

[enlarge]

Figure 1. Cell lysates from WT, APT1-KO and APT2-KO HAP1 cells were analyzed by western blot using an anti-APT1/LYPLA1 antibody. Actin was used a loading control. From [1].

Rabbit monoclonal IgG

Company: Abcam

Antibody: Lysophospholipase 1

Catalog number: ab91606

Summary: Rabbit monoclonal IgG against a synthetic peptide within Lysophospholipase 1 (APT1) (aa 150-250). Reacts with mouse, rat, and human. Suitable for western blot, immunoprecipitation and immunohistochemistry (paraffin). Unsuitable for flow cytometry or immunocytochemistry.

[enlarge]

Figure 2. Immunostaining of endogenous APT1 (green) in WT, APT1-KO and APT2-KO HAP1 cells. From [1].

Western blot | Immunocytochemistry

WT, APT1-KO and APT2-KO HAP1 cells. WB: cells were lysed in RIPA lysis buffer (50 mM Tris, 150 mM NaCl, 0.5% deoxycholate, 0.1% SDS, 1.0% TritonX-100, pH 7.4). IC: Cells were fixed with 3% paraformaldehyde for 20 min at 37 °C.

WB: 5% BSA (BP9703, Fischer) solution in wash buffer (20 mM Tris, pH 7.5, 150 mM NaCl, 0.1% Tween-20) for 1 h.

IC: 0.5% BSA in PBS blocking overnight.

WB: 1:1,000 dilution in 5% BSA solution in wash buffer overnight at 4 °C.

IC: 1:500 dilution.

WB: 1:2,000 dilution goat anti-rabbit IgG HRP (SCBT, sc-2004) in 5% BSA solution in wash buffer for 1h.

IC: Donkey anti-rabbit 568 (Life Technologies, A10042) or Donkey anti-rabbit 647 (Life Technologies, A31573) at room temperature for 30 min.

WB:Super Signal West Pico Plus ( Thermo Scientific, 34577) and recorded on Fluor Chem R (Protein Simple) imaging station.

IC: LSM710 confocal microscope (Zeiss) with a ×63 oil immersion objective (NA 1.4).

The same clone (EPR3667) is sold as Abcam ab91606; EMD Millipore MABS166.