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Synthetic peptide, corresponding to amino acids 1-17 of Human beta Amyloid.
Our Abpromise guarantee covers the use of ab11132 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use at an assay dependent concentration.|
|WB||Use at an assay dependent concentration. Predicted molecular weight: 87 kDa. PubMed: 19955183|
|IHC-P||1/100. Retrieve antigens with 70% Formic acid for 10-30 minutes at room temperature before commencing with IHC staining protocol.|
|IP||Use at an assay dependent concentration.|
ab11132 at 1/100 staining human brain (Alzheimer's diseased) tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step was performed prior to incubation with ab11132 for 16 hours. A biotinylated goat anti-mouse antibody was used as the secondary.
In this experimental set up ab11132 shows clear extracellular and intracellular staining.
ab11132 staining beta amyloid in 15 month TASTPM mouse brain tissue section by Immunohistochemistry (Formalin/PFA fixed paraffin-embedded sections). Tissue underwent fixation in formaldehyde, antigen retrieval in 70% Formic acid buffer for 20 mins and blocking in 1% BSA for 10 mintes at 25°C. The primary antibody was used at 1/100 dilution and incubated with sample for 2 hours. A Biotin conjugated goat polyclonal to mouse IgG1 at 1/200 dilution, was used as secondary.
Immunohistochemistical detection of beta Amyloid using antibody [DE2B4] (ab11132) on transgenic APP23 mouse brain cortex frozen sections. Fixative: Formaldehyde Permeabilization: Yes - 0.1% Tween 20 Blocking step: 1% BSA for 30 mins @ 1 % @ rt°C. Primary Antibody used at 1/200 and incubated for 2 hours in TBS/BSA/Tween 20/azide. Secondary antibody: anti Mouse IgG Alexa Fluor® 594 (1/1000). Immunstaining performed on APP overexpressing TG mouse (APP23) tissue. Unfixed cryosections were fixed in 4% Formalin in PBS before treatment with 70% Formic acid to most effectively reveal antigenic sites. Interestingly, punctate intracytoplasmic positivity is also observed in neurones, using this Ab: this may be APP. Note the fibrillar positivity within the plaque: also that is has a negative centre (older plaques seem to calcify and this is often dislodged).
Image from Skerget K et al, J Biol Chem. 2010 Jan 29;285(5):3201-10. Epub 2009 Dec 2, Fig 8. DOI 10.1074/jbc.M109.024620CHO-K1 cells, expressing either wild-type or Swedish mutant APP, were transfected with the vector encoding stefin B and lysed after 24 hours. The lysates were immunoprecipitated with a polyclonal anti-rabbit stefin B antibody, after which 15% SDS gels and Western blotting with ab11132 at a 1/400 dilution were performed. First and second lanes show results obtained on whole cell lysates of APP-WT and APP-Sw, respectively, and the third and fourth lanes show the corresponding samples after co-immunoprecipitation on anti-stefin B antibody. ab11132 clearly detects a fragment of 15 kDa (according to molecular mass standards) in the APP-WT cells.