TBS or TBS-T, how using respective buffer for primary antibody incubation may change the outcome

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. 



 

TBS or TBS-T buffers for antibody incubation, how can they change the outcome?

 Technical tips

Technical tips

Latest

Can methanol in transfer buffer be replaced by a less toxic alternative?
2025-05-12 Will the precipitation of SDS in the stripping buffer affect antibody stripping efficiency?
2025-04-29 How to check if protein transfer from a gel to a membrane was efficient
2025-04-15 How to detect membrane proteins on Western blot?
2025-03-27 Rinse Right: The Hidden Power of Washing in Western Blots
2025-02-11 Can one Western blot protocol fit all antibodies?
2025-02-07 How to reconstitute lyophilized antibodies?
2025-01-17 What information to look for before purchasing an antibody?
2024-12-30 Membrane activation before protein transfer: which one to use, methanol or ethanol?
2024-12-23 Comparing antibodies between each other, why does it not work?
2024-11-25

Archive


Close
Login / Retailer
Register
Forgot password?