Our Abpromise guarantee covers the use of ab3583 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 a concentration of 5 µg/ml.|
|Inhibition Assay||Use at an assay dependent concentration.|
|ICC||1/50 - 1/200.|
|IHC-Fr||1/50 - 1/200.|
|WB||1/200 - 1/1000. Detects a band of approximately 16 kDa (predicted molecular weight: 16 kDa).|
|Flow Cyt||Use 3-5µl for 106 cells.|
ab3583 at a dilution of 1 / 4000 staining leptin in mouse 3T3-L1 cell lysate by Western blot.
ab3583 staining Leptin in 3T3-L1 cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with 4% paraformaldehyde, permeabilized with 0.1% Triton X-100 and blocked with 1% BSA for 1 hour at room temperature. Samples were incubated with primary antibody (2ug/ml in 0.1% BSA) for 3 hours at room temperature. An Alexa Fluor® 488-conjugated Goat anti-rabbit IgG (H+L) polyclonal was used as the secondary antibody (1/2000) (Panel a). Nuclei were stained with DAPI (Panel b). F-actin stained with Rhodamine Phalloidin (panel c). Merged images shown in panel d.
ab3583 staining Leptin in Human mesenchyml stem cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and blocked with 10% serum for 30 minutes at 22°C. Samples were incubated with primary antibody (1/100 in 10% horse serum) for 1 hour at 22°C. A FITC-conjugated Mouse anti-rabbit IgG1 monoclonal was used as the secondary antibody (1/100).
Immunohistochemical staining of Leptin in Sheep brain using ab3583.
ab3583 staining Leptin in 3T3-L1 cells by Flow Cytometry. The sample was incubated with the primary antibody (3ug/ml in 2.5% BSA) for 2 hours at room temperature. An Alexa Fluor 488®-conjugated Goat anti-rabbit (1/400) was used as the secondary antibody. Red histogram represents ab3583, pink histogram represents isotype control, purple histogram represents unstained control, green histogram represents no primary antibody control