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Our Abpromise guarantee covers the use of ab2539 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/1000 - 1/5000. Predicted molecular weight: 87 kDa.|
|IHC-Fr||Use at an assay dependent concentration.|
|IHC-P||1/100 - 1/200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.|
|ELISA||1/10000 - 1/50000.|
ab2539 at 1/100 staining human brain tissue sections by IHC-P. The sections were formaldehyde fixed and a heat mediated antigen retrieval step was performed prior to incubation with ab2539. A biotinylated goat anti-rabbit IgG was used as the secondary.
In this experimental set up, ab2539 is found to detect intracellular beta amyloid only and is negative for amyloid plaques.
ab2539 staining beta Amyloid in 15 month old TASTPM mouse brain tissue section by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue underwent formaldehyde fixation before heat mediated antigen retrieval in citric acid and then blocking in 1% BSA for 10 minutes in RT was performed. The primary antibody was diluted 1/100 and incubated with sample for 2 hours in the dilution buffer containing TBS, BSA and azide. A Goat anti rabbit IgG was used as a secondary antibody at 1/300 dilution.
Immunohistochemistical detection of beta Amyloid using antibody (ab2539) on TG 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/azide. Secondary Antibody: anti rabbit IgG Alexa Fluor® 488 Dilution 1/1000. Fresh, snap-frozen cryosections of TG APP23 mice were fixed in 4% Formalin in PBS for 20 mins prior to treatment in 70% Formic acid AR (this Ab also works on such sections without acid pretreatment). This section was co-labelled using mouse anti GFAP ( visualising with anti Ms Alexa 594) to illustrate that astrocytes appear to be intimately associated with plaques. Positivity is confined to the extracellular plaques.
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