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Our Abpromise guarantee covers the use of ab37861 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.|
|IHC-P||1/50 - 1/100.|
|Flow Cyt||1/10 - 1/50.
ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.
|IHC-Fr||Use at an assay dependent concentration. PubMed: 24236135|
|WB||1/1000. Detects a band of approximately 97 kDa.|
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast cancer (BC) tissue labelling Axl with ab37861. A peroxidase-conjugated anti-rabbit IgG was used as the secondary antibody, followed by AEC staining.
ab37861 staining Axl in human breast tumor by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with Tween20 and blocked with 10% serum for 30 minutes at 25°C. Samples were incubated with the primary antibody (in PBS + 10% serum) for 12 hours at 4°C. An undiluted Alexa Fluor® 594-conjugated goat anti-rabbit IgG monoclonal was used as the secondary antibody.
ICC/IF image of ab37861 stained SW480 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab37861 at 5µg/ml overnight at +4°C. The secondary antibody (pseudo-colored green) was Alexa Fluor® 488 goat anti- rabbit (ab150081) IgG (H+L) preadsorbed, used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1h at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.
Flow cytometry analysis of WiDr cells labelling Axl (green) with ab37861 compared to a negative control (blue). A FITC-conjugated goat anti-rabbit IgG was used as the secondary antibody.
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