Reagents In The Kit
VIAL 1, Serum Block
1 x 5ml, 5% Normal Serum in 0.01M Phosphate Buffered Saline (PBS) containing 0.1% Sodium Azide (NaN3)
VIAL 2, Reagent Boost
1 x 5ml
VIAL 3, HRP Polymer
1 x 5ml
VIAL 4, Peroxide Blocking Reagent
1 x 5ml, 0.03% Hydrogen Peroxide (H202)
VIAL A, DAB Chromogen
1 x 2ml, 2.5% 3,3’ Diaminobenzidine in stabilizing buffer
VIAL B, DAB Substrate Buffer
4 x 6ml, 0.1% H2O2 in Citric acid buffer
AbD Serotec HISTAR Detection System reagents are designed specifically to replace complex immunohistochemical techniques with a simple kit. This kit provides linking and labeling reagents intended for use with species specific primary antibodies for visualizing cellular antigens in tissue specimens. Visualization is accomplished using an ultra-sensitive indirect labeling method that utilizes novel polymer labeling technology. Secondary antibodies are polymerized directly with Horseradish peroxidase (HRP) into compact polymers bearing a high ratio of enzyme to antibody. This biotin-free system offers enhanced sensitivity and minimal background staining.
All labeling and blocking reagents are pre-diluted for immediate use, and chromogen is provided in concentrated format. This product is suitable for use with the wide range of AbD Serotec ready-to-use or concentrated primary antibody reagents against human targets. Detailed instructions for use can be found in the complete AbD Serotec PDF datasheet, that can be accessed by clicking on the "Download PDF" link above. The procedure has been briefly summarized below:
1) Prepare tissue sections appropriately according to the requirements of the primary antibody being used e.g. apply antigen retrieval if necessary.
2) Incubate paraffin-embedded sections with peroxide blocking reagent for 15 minutes at room temperature. Rinse with Phosphate Buffered Saline (PBS).
3) Apply serum block and incubate for 15 minutes.
4) Apply primary antibody and incubate for 30 minutes.
5) Apply reagent boost and incubate for 20 minutes. Rinse twice with PBS (5 minutes per wash).
6) Apply HRP polymer for 30 minutes, then rinse twice with PBS (5 minutes per wash).
7) Apply DAB substrate solution for 3-5 minutes, then rinse with distilled water.
[Discard unused substrate solution]
8) Counterstain and mount slides using an aqueous mounting medium such as BUF058A