Thermo Scientific Pierce Protein Research Products
SuperBlock T20 (TBS) Blocking Buffer
37536
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I have been working with SuperBlock as a blocking buffer for the last three years and it has dramatically improved the quality of my Western blots and ELISAs. The product is very fast and easy to use. SuperBlock is also biotin free so it is safe to use with my streptavidin ELISAs and Westerns. I consider SuperBlock the best blocker on the market for protocols which require the use of streptavidin.
ELISA
Biotin labeled proteins
Biotin labeled proteins were coated on ELISA plate for 1 hour at 37 degrees Celsius. Product was blocked overnight using 250ul of SuperBlock T20 (TBS) Blocking Buffer. Wells were washed four times using TBST. Primary antibodies were diluted in 1mL SuperBlock T20 (TBS)+ 4mL TBST. 200ul of antibody mixture were added to each well and incubated at 37 degrees Celsius for 1 hour. Wells were washed again four times using TBSTSecondary antibodies and streptavidin alkaline phosphatase (to ensure protein coating) were diluted in 1mL SuperBlock T20(TBS)+4mL TBST. Plate was incubated at room temperature for 1 hour. Plate was washed 4 times with TBST. Plate was developed using CDP-Star Stabilized Alkaline phosphatase substrate
If your background remains high make sure to shake the SuperBlock. It contains colloidal carbon which needs to be resuspended.
Very positive values were observed in the wells which contained biotin labeled proteins. Very low readings where the non biotin labeled proteins were added when probed with streptavidin.
NA
Easy to use
Incubation times may need to be extended
This product is very easy to use and is a wonderful product if you need a biotin free blocker.
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