48 h after transfection, proteins of the cell were extracted. Protein of the clinical sample and the cultured cell were separated by SDS-PAGE and transferred to nitrocellulose membranes, which were blocked with 5% fat-free milk for 1 h. Then, the membranes incubated with anti-APP antibody (1:1,000; Abcam, Cambridge, UK) at 4°C overnight. Anti-β-actin antibody was served as the internal reference. The membranes were incubated with secondary antibodies for 30 min at room temperature after washing thoroughly. We detected the results by enhanced chemiluminescence technique (Amersham, Piscataway, NJ, USA) and quantified the level of expression of these proteins by application of Image J software.
Anti app antibody
Manufactured by Abcam
Sourced in United Kingdom
Anti-APP antibody is a laboratory reagent used to detect the presence and expression levels of the amyloid precursor protein (APP) in biological samples. This antibody recognizes the APP protein and can be used in various research applications such as Western blotting, immunohistochemistry, and ELISA.
Automatically generated - may contain errors
Lab products found in correlation
Enhanced chemiluminescence technique, by Cytiva (1 mentions)
Lysis buffer, by Cell Signaling Technology (1 mentions)
Horseradish peroxidase linked anti rabbit igg, by Cell Signaling Technology (1 mentions)
Anti bace1 antibody, by Abcam (1 mentions)
Anti adam9, by Cell Signaling Technology (1 mentions)
Trypsin edta, by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Zeocin, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by American Type Culture Collection (1 mentions)
Rabbit anti gapdh, by Cell Signaling Technology (1 mentions)
Aperio slide scanner, by Leica (1 mentions)
3 protocols using anti app antibody
1
Western Blot Analysis of APP
2
APP Processing Regulation Protocol
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Rabbit anti-APP antibodies to detection the C-terminal of APP were purchased from Sigma-Aldrich Co (St. Louis, MO, USA). FBS was purchased from ATCC Company (Manassas, VA, USA). DMEM, penicillin/streptomycin, G418, and 0.25% trypsin-EDTA were purchased from GIBCO–BRL Company (Carlsbad, CA, USA). Zeocin were purchased from Invitrogen Company (Carlsbad, CA, USA). Rabbit anti-GAPDH, anti-rabbit horseradish peroxidase linked IgG, anti-ADAM9 antibodies and lysis buffer were obtained from Cell Signaling Technology (Danvers, MA, USA). Anti-BACE1 antibody, and anti-APP antibody to detection both mAPP and imAPP were obtained from Abcam Company (Cambridge, UK). Anti-ADAM10 antibody was obtained from Calbiochem Company (San Diego, CA, USA). Anti-TACE and anti-ADAM17 antibodies were obtained from Chemicon Company (Billerica, MA, USA). All other chemicals were of analytical grade obtained from Sigma-Aldrich Co.
3
Immunohistochemical Analysis of APP in FFPE Tissue
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The method of choice for antigen retrieval in immunohistochemistry was by heating the FFPE sections bathed in TE buffer (10 mmol/L Tris, pH 8.0, and 5 mmol/L EDTA) in a pressure cooker. The sections were then incubated with anti-APP antibody (catalog number ab32136; Abcam, Cambridge, UK) (dilution, 1:250) at 4 C overnight, and the signal was visualized using the EnVisionþ Dual Link System (Agilent, Santa Clara, CA). 19 Negative control was prepared by replacing the primary antibody with phosphatebuffered saline. 20 Fetal and placental tissue found to be positive was used as positive control. The stained sections were scanned with an Aperio slide scanner (Leica Biosystems, Wetzlar, Germany). The representative regions of trophoblast on each section were marked by a pathologist (A.N.Y.C.), and the staining intensity of the regions was evaluated by the positive pixel count v9 algorithm included in the system.
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