Protein Detector™ Microarray Dot Blot Kits offer high sensitivity, accuracy and quantitation over a wide range in low-density protein array (96- and 384-spot formats) detection. They are ideal for performing reverse-phase protein arrays and for profiling hundreds of samples for the presence of low abundant antigens. Protein Detector kits are based on a biotin/streptavidin system and provide substantial amplification over enzyme-labeled antibody methods commonly used to detect Western blots.
In addition, the strong and specific interaction between biotin and streptavidin permits the use of stringent incubation and washing conditions, thus making it a highly robust method for microarray studies. KPL's blocking reagent included in the kit is formulated for microarray blotting and delivers a comparatively higher signal to noise ratio that traditional blockers.

KPL offers two complete kits with a choice of colorimetric or chemluminescent substrate, matched blocker and protocols for scientists who want the convenience of an optimized kit. Figure 1 below demonstrates that both the AP colorimetric kit and the AP chemiluuminescent kit can be used to detect as little as 30 pg of spotted IgG prin a reverse phase protein microarray. All reagents necessary for detection are included in the kits. Users supply the microarray membrane, experimental samples and primary antibodies.

Figure 1:  Mouse IgG samples were spotted onto nitrocellulose membranes and detected using the indicated kit according to the kit product manual.  The amounts of protein spotted (both panels) are as follows (higher concentrations toward the left and lower concentrations toward the right):  Row 1, two-fold dilutions starting at 2 ng and ending at 16 ng; Row 2, two-fold dilutions starting at 7.8 ng and ending at 61 pg; Row 3, two-fold dilutions starting at 30 pg and ending at 0.24 pg.  Panel A— AP colorimetric kit; Panel B—AP chemiluminescent kit.

Kit Components

1. Biotinylated Goat Anti-Rabbit IgG (H + L)
2. Biotinylated Goat Anti-Mouse IgG (H + L), human serum adsorbed
3. Reconstitution Buffer
4. AP-labeled Streptavidin
5. 20X Wash Solution Concentrate
6. Microarray Blocking Powder
7. Microarray Blocking Solution
8. Microarray AP Chemiluminescent Substrate (KP-56-12-50) or BCIP/NBT Substrate (KP-56-11-50)