Ferangi Blue™ Chromogen KitChromogen Kit

ISO9001:2000

CERTIFIEDControl Number: 901-FB812-101910

Intended Use:For In Vitro Diagnostic Use

Summary & Explanation:When in the presence of alkaline phosphatase (AP) enzyme, Ferangi Blue™ produces abright royal blue precipitate that is insoluble in most organic solvents (except xylene ortoluene) and can be coverslipped with a permanent mounting media. This product comes in a stable four-component system, consisting of liquid Ferangi Blue™ Substrate, Ferangi Blue™ Chromogen, Ferangi Blue™ Buffer and Ferangi Blue™ Levamisole. When using an alkaline phosphatase system, tris buffer (pH 7.6) should beused as a rinsing buffer. PBS should never be used! Phosphates act as a competitive inhibitor to alkaline phosphatase enzymes. For optimum performance, use ImmunocareTBS Wash Buffer with AP activator.

Known Applications:Immunohistochemistry (formalin-fixed paraffin-embedded tissues)

Catalog Number: FB812 S

Description: 100 ml

Supplied As:1. Ferangi Blue™ Substrate (FB812SB) 1.4 ml 2. Ferangi Blue™ Chromogen FB812CH) 1.4ml3. Ferangi Blue™ Levamisole (FB812LV) 3ml 4. Ferangi Blue™ Buffer (FB812BF) 100 ml5. Mixing vial (VL103) 1ea6. Dropper Bottle (DB812) 1ea

Storage and Stability:Store at 2ºC to 8ºC. Do not use after expiration date printed on vial. If reagents are stored under conditions other than those specified in the package insert, they must be verified by the user. Diluted reagents should be used promptly; any remaining reagentshould be stored at 2ºC to 8ºC.

Protocol Recommendations:Add 1 drop (32µl) of Ferangi Blue Substrate (A) to 2.5 ml of Ferangi Blue Buffer. Mixwell. Then add 1 drop of Ferangi Blue Chromogen (B) to the mixture. Mix well. Make just prior to use. The Ferangi Blue mixture is stable for 20-30 minutes. After using mixing vial and/or dropper bottle, rinse with 70% alcohol and then wash in several changes of deionized water.

Materials and Reagents Needed But Not Provided:Microscope slides, positively chargedDesert chamber* (Drying oven)Positive and negative tissue controlsXylene (Could be substituted with xylene substitute*)Ethanol or reagent alcoholDecloaking chamber* (Pressure cooker)Deionized or distilled waterWash buffer*(TBS)Pretreatment Reagents*Enzyme Digestion*Avidin-Biotin Blocking Kit*(Labeled Streptavidin Kits Only)Peroxidase block*Protein block*Primary antibody*Negative Control Reagents*Detection Kits*Detection Components*Hematoxylin*Bluing Reagent*Mounting medium*

* Biocare Medical Products: Refer to a Biocare Medical catalog for further informationregarding catalog numbers and ordering information. Certain reagents listed above arebased on specific application and detection system used.

Performance Characteristics:The protocols for a specific application can vary. These include, but are not limited to:fixation, heat-retrieval method, incubation times, tissue section thickness and detectionkit used. Due to the superior sensitivity of these unique reagents, the recommended incubation times and titers listed are not applicable to other detection systems, as results may vary. The data sheet recommendations and protocols are based on exclusive use of Biocare products. Ultimately, it is the responsibility of the investigator to determine optimal conditions. These products are tools that can be usedfor interpretation of morphological findings in conjunction with other diagnostic tests and pertinent clinical data by a qualified pathologist.

Precautions:This product is not classified as hazardous. The preservative used in this reagent is Proclin 950 and the concentration is less than 0.25%. Overexposure to Proclin 950 or chromogen can cause skin and eye irritation and irritation to mucous membranes and upper respiratory tract. The concentration of Proclin 950 in this product does not meet the OSHA criteria for a hazardous substance. Wear disposable gloves when handling reagents.Specimens, before and after fixation, and all materials exposed to them should be handled as if capable of transmitting infection and disposed of with proper precautions.Never pipette reagents by mouth and avoid contacting the skin and mucous membraneswith reagents and specimens. If reagents or specimens come in contact with sensitive areas, wash with copious amounts of water. Microbial contamination of reagents may result in an increase in nonspecific staining. Incubation times or temperatures other than those specified may give erroneous results. The user must validate any such change. The MSDS is available upon request.Consult OSHA, federal, state or local regulations for disposal of any toxic substances.ProclinTM is a trademark of Rohm and Haas Company, or of its subsidiaries or affiliates.Warnings:1. This chromogen solution contains Methanol.2. Wear appropriate personal protective equipment and clothing.3. May cause dermatitis. Avoid contact with skin and eyes.4. Harmful if swallowed.5. The MSDS is available upon request.

Quality Control:Refer to NCCLS Quality Assurance for Immunocytochemistry approved guidelines, December 1999 MM4-A Vol.19 No.26 for more information on tissue controls.

Protocol Recommendations cont'd:Optional:Use only if endogenous alkaline phosphatase staining is a problem. Add 1 drop of Ferangi Blue Substrate (A) and 1-2 drops of Ferangi Blue Levamisole solution (C) to 2.5 ml of Ferangi Blue Buffer. Mix well. Then add 1 drop of Ferangi Blue Chromogen (B) to the solution and mix well. Solution may turn slightly turbid.1. Rinse tissue with TBS Wash Buffer.2. Apply the Ferangi Blue mixture to the tissue section. Incubate 5 minutes.3. Rinse tissue with deionized water (D.I.).4. Counterstain with Weigert’s hematoxylin for 5 to 10 seconds or Nuclear Fast Red for 10 minutes. Rinse in D.I. water.5. Dehydrate in 3 changes of 100% alcohol and clear in 3 changes in a xylene substitute (Biocare’s Slide Brite).6. Mount and coverslip with non-xylene permanent mounting medium.Protocol Notes:1. Ferangi Blue can be used with DAB, Bajoran Purple and Fast Red for double and triple stain procedures.2. Hydrogen peroxide block does not inhibit AP staining. In some cases, a hydrogen peroxide block can produce higher contrast and cleaner sections. It does not hurt the AP procedure.Ferangi Blue is partially soluble in xylene. Avoid xylene and xylene-based coverslipping mounting media. Also, avoid prolonged dehydration steps. Mount in a non-xylene based mounting media (Cover*Safe, American Master Tech, 800-860-4073).

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Ferangi Blue™ Chromogen KitChromogen Kit

ISO9001:2000

CERTIFIEDControl Number: 901-FB812-101910

Limitations & Warranty:There are no warranties, expressed or implied, which extend beyond this description. Biocare is not liable for property damage, personal injury, or economic loss caused by this product.

Troubleshooting:Follow the antibody specific protocol recommendations according to data sheet provided. If atypical results occur, contact Biocare's Technical Support at1-800-542-2002.

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