Recombinant human Lymphocyte Activation Gene 3 (soluble form).
Our Abpromise guarantee covers the use of ab40465 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|Flow Cyt||Use 1µg for 106 cells.|
|IHC-P||Use at an assay dependent concentration.|
|IP||Use a concentration of 10 µg/ml.|
|WB||Use a concentration of 5 µg/ml. Predicted molecular weight: 58 kDa.|
|ELISA||Use a concentration of 5 µg/ml.|
|Blocking||Use at an assay dependent concentration.|
ab40465 staining Lymphocyte Activation Gene 3 in Human submandibular gland (disease) tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with blocking buffer for 5 minutes at 24°C; antigen retrieval was by heat mediation in a sodium citrate buffer, pH 6.0. Samples were incubated with primary antibody (1/100 in PBS) for 2 hours at 25°C. An undiluted Biotin-conjugated anti-mouse IgG monoclonal was used as the secondary antibody.
Overlay histogram showing Jurkat cells stained with ab40465 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab40465, 1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.