Introduction
Skim Milk Powder can be used to prepare blocking buffer, primary antibody diluent and secondary antibody diluent in Western detection. Skim Milk Powder is one of the most commonly used blocking buffer components for Western detection.
This product has good solubility. After blocking, it can effectively reduce the non-specific combination of primary antibody or secondary antibody and membrane, reduce the background and enhance the signal-to-noise ratio.
The blocking buffer prepared by Skim Milk Powder is different from that prepared by BSA, Skim Milk Powder is opaque, and BSA is transparent.
For Western blotting detection of specific target protein, sometimes BSA is better as the main blocking component, sometimes Skim Milk Powder is better. It’s according to the antigen and antibody specificity.
Components
|
Cat
|
Products
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5
g
|
10
g
|
50
g
|
Storage
|
|
E-BC-R337
|
Skim Milk Powder
|
5 g
|
10 g
|
50 g
|
RT
|
Experimental Procedure
1. Preparation of Western blotting blocking buffer. The concentration of Skim Milk Powder is usually 3~5% (w / V). It is recommended to use Elabscience® 10× TBST (E-BC-R335) and dilute it to 1 × TBST working buffer with deionized water at the rate of 1:9 as dilution buffer.
2. The primary and secondary antibodies are recommended to be diluted with the blocking buffer.
3. Wash the membrane for 1~2 min after the sample transfer. Immerse the membrane in the Western blotting blocking buffer and block at RT for 60 min.
4. Wash the membrane for 1~2 min, 1~2 times. Then incubate the primary antibody and secondary antibody which are also dilute with the blocking buffer.
Storage
Store at RT. Valid for 12 months.
Cautions
1. Usually we block the membrane at RT for 60 min. If the antibodies are with very high background, please try to block at 4°C overnight.
2. The components of Skim Milk Powder are complex and may contain trace biotin, so it is not suitable for biotin-avidin detection system.
3. This product is not suitable for anti-Goat or anti-Sheep secondary antibodies.
4. There is no blocking buffer suitable for all experimental systems, for some special experiments, it is necessary to consider using other more suitable blocking buffer according to the specific situation.
5. Please keep the Western blotting membrane or immunohistochemical slice moist, otherwise it may produce abnormal background.
6.This product is for research use only. It couldn’t be used for clinical diagnosis or treatment, food or medicine, and can’t be stored in residence.
7.For your safety and health, please wear the lab coat and disposable gloves before the experiments.
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