This is a knockout-validated antibody summary, based on the publication "Region-Specific Differences in Amyloid Precursor Protein Expression in the Mouse Hippocampus", 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. Immunofluorescence APP in hippocampal sections of adult wild type and APP deficient mice. (A) Dorsal hippocampus of wild type (+/+) mice. The dentate gyrus (DG) shows only a weak signal, whereas a more intense labeling is seen in Ammon’s horn (CA1–3). (B,C) Immunofluorescence is detectable in the granule cell layer (gcl) and molecular layer (ml) of the DG in wild type but not in brain sections of APP deficient mice.

Rabbit monoclonal IgG

Company: Epitomics (now Abcam)

Antibody: Amyloid Precursor Protein

Catalog number: ab32136

Summary: Rabbit monoclonal IgG against a synthetic peptide within human Amyloid Precursor Protein (aa 750 to the C-terminus). Reacts with mouse, rat and human. Suitable for western blot, immunocytochemistry/immunofluorescence, immunohistochemistry (paraffin, frozen and PFA perfusion fixed frozen), immunoprecipitation and flow cytometry.

Immunohistochemistry | Western blot

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Figure 2. Western blot analysis of APP protein in adult mouse hippocampus shows the typical set of bands corresponding to holoAPP (hAPP) protein (~95–100 kDa) in adult wild type (APP+/+) mice but not in APP deficient (APP−/−) tissue. From [1].

Dorsal hippocampus of wild type (+/+) and APP deficient (APP−/−) mice.

IHC: Mice were transcardially perfused with 0.9% sodium chloride (NaCl) followed by 4% paraformaldehyde (PFA) in phosphate-buffered saline (pH 7.4) and brains were removed, post-fixed for 4–24 h in 4% PFA and sectioned in the coronal plane (40 μm) using a vibratome (VT1000 S, Leica Microsystems).

WB: Tissue was homogenized in 10× volume of homogenization buffer (20 mM Tris, 500 mM NaCl, 0.5% CHAPS, 5 mM EDTA).

IHC: Free-floating sections were incubated in a blocking buffer containing 0.5% Triton X-100 and 5% bovine serum albumin (BSA) in 0.05 M Tris-buffered saline (TBS) for 30 min at room temperature.

WB: Odyssey Blocking Buffer (LI-COR Biosciences) at room temperature for 60–120 min.

IHC: diluted in 0.1% Triton X-100 and 1% BSA in 0.05 M TBS overnight at 4°C.

WB: diluted in 1:1 Odyssey Blocking Buffer with TBS and 0.1% Tween20 overnight at 4°C.

IHC: 1:2,000 dilution secondary Alexa-conjugated antibodies (Invitrogen, Waltham, MA USA) for several hours at room temperature.

WB: IRDye800CW conjugated secondary antibody (LI-COR Biosciences) at room temperature for 45 min.

IHC: Fluorescent images were acquired using a digital camera (Digital Sight DS-M5c, Nikon, Germany) or confocal microscopy (Eclipse C1 Plus, Nikon).

WB: Odyssey Infrared Imaging System (LI-COR Biosciences).