- Intended Use
- Bio-Rad 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.
The HISTAR Detection System is designed for the staining of Human tissue sections. This kit is suitable for use with mouse and rabbit primary antibodies only.
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 Bio-Rad ready-to-use or concentrated primary antibody reagents against human targets. Detailed instructions for use can be found in the complete Bio-Rad PDF datasheet, that can be accessed by clicking on the "Instructions For Use" link. 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.
- 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, 3% 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
- Additional Reagents Required
- 1) Primary antibody and positive control
2) Phosphate Buffered Saline (PBS), pH7.4 (+ 0.2)
4) Mounting media
Bio-Rad can supply a number of these products separately, please see the ‘Recommended Useful Reagents’ section.
- Store at +4oC. DO NOT FREEZE.
This product should be stored undiluted.
- Guaranteed until date of expiry. Please see product label.
- For research purposes only
This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.
Applications of Histar Detection Kit
|Immunohistology - Frozen
||Ready to use
|Immunohistology - Paraffin
||Ready to use
Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
- Instructions For Use
- Instructions for use can be found at www.bio-rad-antibodies.com/uploads/IFU/STAR3000A.pdf
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Useful Reagents Available