Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Claudin-1 (D5H1D) XP® Rabbit mAb #13255 (brown) followed by a counterstain with Hematoxylin (blue).
Chemical structure of Hematoxylin.
Hematoxylin is a blue nuclear counterstain for use after the completion of immunohistochemical assays. It yields crisp nuclear staining with superior contrast with SignalStain® DAB Substrate Kit #8059 and is also compatible with SignalStain® Mounting Medium #14177.
A) Solutions and Reagents
1. Hematoxylin #14166
Additional Reagents (Not Supplied)
1. Acid Rinse Solution: To prepare 100ml, add 2ml of glacial acetic acid to 98 ml of dH20.
2. Bluing Solution: To prepare 100ml, add 1.5 ml of 30% ammonium hydroxide (NH4OH) to 98.5ml of 70% ethanol
3. SignalStain® Mounting Medium #14177
B) Nuclear Counterstaining
1. Rinse slides in tap water.
2. Completely submerse slides in Hematoxylin #14166 and incubate for 1-5 minutes.
3. Wash slides with tap water until rinse water is clear.
4. Immerse slides 10 times in acid rinse solution.
5. Rinse slides 10 times in tap water.
6. Immerse slides for 1 minute in bluing solution
7. Rinse slides 10 times in tap water.
8. Mount sections with SignalStain® Mounting Medium #14177.
Supplied as a working solution. Store at room temperature. Product is stable for 12 months upon receipt when stored properly.
Hematoxylin is a blue nuclear counterstain for use in immunohistochemical assays. It yields crisp staining detail with superior contrast when used in conjunction with SignalStain® DAB Substrate Kit #8059. It is also compatible with SignalStain® Mounting Medium #14177.
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