Immunohistochemical analysis of paraffin-embedded human colon carcinoma using APP (E3F3P) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human endometrioid adenocarcinoma using APP (E3F3P) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded normal human brain using APP (E3F3P) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human ovarian serous carcinoma using APP (E3F3P) Rabbit mAb (left) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).
Immunohistochemical analysis of paraffin-embedded A549 cell pellet, control shRNA-treated (left) or APP shRNA-treated (right), using APP (E3F3P) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human spleen (left) or adenoid cystic carcinoma of the trachea (right) using APP (E3F3P) Rabbit mAb (top) or APP (E8B3O) XP® Rabbit mAb #29765 (bottom). These two antibodies detect independent, unique epitopes on human APP. The similar staining patterns obtained with both antibodies help to confirm the specificity of the staining.
Immunohistochemical analysis of paraffin-embedded mouse brain using APP (E3F3P) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded mouse lung using APP (E3F3P) Rabbit mAb.
|MW (kDa)||9, 100-140|
|Immunohistochemistry (Paraffin)||1:50 - 1:200|
Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
NOTE: Do not allow slides to dry at any time during this procedure.
For Citrate: Heat slides in a microwave submersed in 1X citrate unmasking solution until boiling is initiated; follow with 10 min at a sub-boiling temperature (95°-98°C). Cool slides on bench top for 30 min.
|SignalStain® Boost IHC Detection Reagent (HRP, Rabbit) #8114||SignalStain® Boost IHC Detection Reagent (AP, Rabbit) #18653|
|SignalStain® DAB Substrate Kit #8059||SignalStain® Vibrant Red Alkaline Phosphatase Substrate Kit #76713|
|SignalStain® Vivid Purple Peroxidase Substrate Kit #96632|
NOTE: Use of detection reagents other than those specified in this protocol may require further optimization of the primary antibody to account for the different sensitivities of the detection reagents.
posted February 2010
revised June 2020
Protocol Id: 283
APP (E3F3P) Rabbit mAb recognizes endogenous levels of total APP protein.
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human APP protein.
Amyloid β (Aβ) precursor protein (APP) is a 100-140 kDa transmembrane glycoprotein that exists as several isoforms (1). The amino acid sequence of APP contains the amyloid domain, which can be released by a two-step proteolytic cleavage (1). The extracellular deposition and accumulation of the released Aβ fragments form the main components of amyloid plaques in Alzheimer's disease (1). APP can be phosphorylated at several sites, which may affect the proteolytic processing and secretion of this protein (2-5). Phosphorylation at Thr668 (a position corresponding to the APP695 isoform) by cyclin-dependent kinase is cell-cycle dependent and peaks during G2/M phase (4). APP phosphorylated at Thr668 exists in adult rat brain and correlates with cultured neuronal differentiation (5,6).
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