Bio-Rad eBook - 2019 - 7

Blocking Reagents

Following transfer, before immunodetection, unoccupied antibody binding sites on the membrane must be blocked
to prevent nonspecific antibody binding. Failure to completely block these sites can lead to high backgrounds that
obscure the signal.
The best blocker for each experiment will depend on the antibody and membrane type. Optimize the detection
system for maximal signal with lowest background by testing several blocking agents. Modern formulations like
Bio-Rad's EveryBlot Blocking Buffer are universal, performing well across a wide range of targets, sample types,
and detection methods.

EveryBlot Universal Blocking Buffer

EveryBlot Blocking Buffer provides 5 minute blocking and maximum
sensitivity for all western blots regardless of detection method.

*	 Reduces nonspecific antibody binding to reduce background while
maintaining excellent sensitivity
*	 Compatible with both chemiluminescent and fluorescent detection
*	 Contains no phosphate-based buffers. Ideal for use with phosphospecific antibodies
*	 Complete blocking in 5 minutes

See the Data
EveryBlot Universal Blocking Buffer outperforms other blocking buffers in all western blotting applications:
Chemiluminescent Detection

Fluorescent Detection
Other vendor's new
fluorescence-optimized	
blocker, 1 hour

EveryBlot	Blocking	Buffer,	
5 minutes

Milk,
30 minutes

EveryBlot	Buffer,	
5 minutes
RNA Helicase A

RNA Helicase A (DyLight 800)
ALIX (StarBright 700)

p53

Tubulin (hFAB Rhodamine)

Low Background across the Entire Spectrum

PRDX3

Phosphoprotein Detection
Calyculin A

EGF

EGF

Total AKT2
AKT2 pSer474

Total EGFR
EGFR pTyr845

Total EGFR
EGFR PSER1047

EveryBlot 5 Minute Universal Blocking Buffer
bio-rad.com/EveryBlot

7

https://www.bio-rad.com/EveryBlot https://www.bio-rad.com/EveryBlot https://www.bio-rad.com/EveryBlot https://www.bio-rad.com/EveryBlot

Bio-Rad eBook - 2019

Table of Contents for the Digital Edition of Bio-Rad eBook - 2019