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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 concentration.|
|IHC-P||Use a concentration of 10 µg/ml.
Proteinase K pre-treatment (starting treatment time 3 min) is recommended for antigen retrieval. Use 20 ug/ml in TE buffer, pH 8.0.
|RIA||Use at an assay dependent concentration.|
|IHC-Fr||Use a concentration of 10 µg/ml.|
|Flow Cyt||Use at an assay dependent concentration. Ab18536-Rat monoclonal IgG2b, is suitable for use as an isotype control with this antibody.|
|ICC||Use at an assay dependent concentration.|
|IHC-FoFr||Use a concentration of 10 µg/ml.|
|ICC/IF||Use at an assay dependent concentration.|
|IHC-R||Use a concentration of 10 µg/ml.|
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).
ab6640 staining F4/80 in mouse spleen tissue sections by Immunohistochemistry (PFA perfusion fixed frozen sections). Tissue samples were fixed by perfusion with paraformaldehyde and blocked with 5% serum for 45 minutes at 21°C. The sample was incubated with primary antibody (1/100 in PBS + 5% FBS) at 4°C for 16 hours. An Alexa Fluor® 488-conjugated goat anti-rat IgG (H+L) polyclonal (1/200) was used as the secondary antibody.
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.
This image is courtesy of an Abreview submitted by Pawel MazurBlocking Step: 10% Milk for 1 hour at room temperature
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"