| In this experiment, the following conditions were applied: Total protein load/well: 30 µg Blocking: 1 h, 5% non-fat milk Primary antibody: 1:1000 O/N 4°C with TBS-T or TBS buffers Secondary antibody: Goat anti-rabbit, 1:25 000 1hr RT with TBS (AS09 602, Agrisera) Detection: Agrisera ECLSuperBright, 5 min As seen in the image to the righ, using TBS buffer without Tween 20 resulted in the detection of the target protein in the overexpressing line and the wild type. |
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Are monoclonal antibodies better, compare to polyclonal?2023-11-30 Why, to detect some proteins, 6M urea must be included in the extraction and loading buffers?
2023-09-01 I have not used this antibody before, what conditions of Western blot do you recommend?
2023-07-27 Which subclasses of immunoglobulin G are found in rabbit serum?
2023-07-18 Antibody suddenly does not work any longer, what can be the reason?
2023-07-11 Can an antibody made to a full-length protein distinguish between 2 isoforms?
2023-05-10 What is a reason for so called "ghost bands" to appear on blots?
2023-03-30 One of the ways to remove background on a Western blot
2023-03-24 How to predict in which technique a given antibody may work?
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