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Thioglycollate stimulated peritoneal macrophages from C57/BL mice
Our Abpromise guarantee covers the use of ab6640 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use a concentration of 10 µg/ml. Use under non reducing condition. Detects a band of approximately 160 kDa (predicted molecular weight: 102 kDa). (predicted molecular weight of precursor protein: 102 kDa; protein is heavily glycosylated). Block in 5% milk for 1 hour.|
|IP||Use at an assay dependent dilution.|
|IHC-P||Use a concentration of 10 µg/ml. Perform heat mediated antigen retrieval (e.g. with citrate buffer) or carry out Proteinase K pre-treatment (starting treatment time 3min).|
|RIA||Use at an assay dependent dilution.|
|IHC-Fr||Use a concentration of 10 µg/ml.|
|Flow Cyt||Use at an assay dependent dilution.|
|ICC||Use at an assay dependent concentration.|
|IHC-FoFr||Use a concentration of 10 µg/ml.|
|ICC/IF||Use at an assay dependent dilution.|
|IHC-R||Use a concentration of 10 µg/ml.|
ab6640 staining F4/80 (red) in Mouse intestine tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with acetone and blocked with 10% serum for 1 hour at 21°C. Samples were incubated with primary antibody (1/10µg/ml in PBS + 10% serum) for 16 hours at 4°C. An Alexa Fluor® 555-conjugated Donkey anti-rat IgG polyclonal (1/1000) was used as the secondary antibody. Blue - nuclei.
ab6640 staining mouse brain tissue sections by IHC-FoFr. Sections were PFA fixed and blocked with 0.5% TNB for 30 minutes at 25°C. The primary antibody was diluted 1/10 and incubated with the sample for 18 hours at 4°C. A biotinylated goat anti-rat antibody, diluted 1/500, was used as the secondary. This image demonstrates quiescent microglia in the normal mouse brain (red).
ab6640 staining F4/80 in mouse adipose tissue section by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue underwent formaldehyde fixation and blocking with 5% serum was performed for 20 minutes at 20°C. The primary antibody was diluted 1/150 and incubated with sample in Tris + 5 % normal rabbit serum for 1 hour at 20°C. A Biotin conjugated rabbit polyclonal to rat IgG was used at dilution at 1/50 as secondary antibody. The antibody stained interstitial macrophages in the adipose tissue of lean mice (A), and macrophage associated with "crown-like structure" in mice fed high-fat diet (B).
This image is courtesy of an Abreview submitted by Pawel MazurBlocking Step: 10% Milk for 1 hour at room temperature
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