Full length native protein (purified) (Pig).
Our Abpromise guarantee covers the use of ab769 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|IHC-P||Use a concentration of 10 µg/ml.|
|ELISA||Use at an assay dependent concentration.|
|RIA||Use at an assay dependent concentration.|
|WB||Use at an assay dependent concentration.
Works best under non reducing conditions.
|Sandwich ELISA||Use a concentration of 1 µg/ml. For sandwich ELISA, use this antibody as Capture at 1µg/ml with ab9538 as Detection.|
|ICC||Use at an assay dependent concentration.|
|IP||Use at an assay dependent concentration.|
|Functional Studies||Use at an assay dependent concentration. Effectively blocks the transferrin-transferrin receptor interactions.|
The sample (commercial human transferrin) (1µg/ml in 1x PBS) was mixed with 4x loading buffer (non reducing) or with Laemmli buffer (reducing) and warmed for 3 minutes.
Volume of sample:5; 3; 1 µl of sample + 3; 2; 2 µl of buffer
SDS Page: 12% AA-gel, U=180V, 90 minutes, volume of loading sample:8; 5;3 µl
Marker – 3 µl
WB: Nitrocelulose membrane (8x5.5 cm), I=35mA, 90minutes
Blocking solution: 1xPBS/5% BSA, 4°C, overnight
Primary Ab: membrane was incubated in 5 ml DMEM/10%FTS + 10 µl HTF-14 (primary Ab) (60 minutes, room temperature)
Washing solution: 1x PBS/0.1% Tween
Secondary Ab: IRDye®800CW Goat anti-mouse IgG1-specific, dilution 1/5000, diluting solution – 5% BSA/1x PBS, incubation for 60 minutes, low temperature.
Detection: Li-COR odyssey
No reducing agent should be added to the sample buffer. The sample can be denatured and processed by common Western blotting protocols, but the disulfide bonds must not be reduced.