AdvanBlock-Chemi blocking solution

AdvanBlock™-Chemi is a novel blocking solution optimized to enhance specific antibody-antigen interactions for the best results in chemiluminescent Western Blots. This all-in-one blocking and antibody incubation solution is designed to improve sensitivity and decrease overall background. Non-specific binding caused by low quality antibodies is reduced while signal from the specific antibody-antigen complex is stabilized and enhanced.

AdvanBlock™-Chemi is provided as a convenient ready-to-use 1x solution intended to directly replace other commonly used blocking buffers for Western Blotting.

AdvanBlock™-Chemi provides high sensitivity and the lowest background.

Figure 1. The appearance of non-specific bands on the Western blot is reduced with AdvanBlock™-chemi, while the specific signal is enhanced compared to three traditional buffers. HeLa whole cell lysates were subjected to Western blotting with an anti-phospho-STAT3 antibody.

Reuse your antibody up to 4 times when diluted in AdvanBlock™-Chemi

Figure 2. TAnti-phospho STAT3 demonstrates increased stability in AdvanBlock™-Chemi compared to 5% Milk-PBST + 0.05% NaN3. (A) 5-fold serial dilutions of IFNα-treated HeLa lysate (5, 1 and 0.2 μg) were separated by SDS-PAGE and transferred to a nitrocellulose membrane. The blots were blocked with AdvanBlock™-Chemi or 5% Milk PBST before incubation with anti-phospho STAT-3 (Cell Signaling Technology #9145S). Sodium azide was added to the milk solution used to dilute the primary antibody to prevent microbial growth. After the primary antibody incubation step was complete, each blocking solution was transferred to a sterile 15 mL conical tube and stored at 4°C. Signal was detected with WesternBright® ECL. In addition to the enhancement of specific signal, the antibody diluted in AdvanBlock™-Chemi shows increased stability demonstrated by the reuse of the antibody solution more than 4 times over the course of two weeks.

AdvanBlock™-Chemi enhanced the Western blot signal compared to non-fat dry milk with 20 common antibodies and different sample types

Table 1: Twenty different antibodies were used in Western blotting using NFDM or AdvanBlock™-chemi.  Background corrected mean signal for each band was measured and then normalized to the background corrected mean signal of NFDM.  The median fold-increase overall was 4.1.

Figure 3:   HeLa cells were treated with IFNα and the whole cell lysates were resolved and probed by Western blotting for STAT1.  Increasing quantities of lysates were loaded on the gel, Western blotted using either milk or AdvanBlock-Chemi (3a, 3b) and then graphed (figure 3c).

Figure 4. Western blotting against ERK-2, pStat-1, and Tubulin shows stronger signals when AdvanBlock-Chemi is used compared to Milk.

• AdvanBlock-Chemi brochure
• AdvanBlock-Chemi Paper, Electrophoresis 2017
• AdvanBlock-Chemi short protocol
• AdvanBlock-Chemi SDS