Exalpha Biologicals, Inc.

	ApoMark™ DNA Fragmentation Apoptosis Detection Kit
The Exalpha Biologicals, Inc. ApoMark™ DNA Fragmentation Detection Kit is a non-isotopic system for the labeling of DNA breaks in apoptotic cell nuclei in paraffin-embedded tissue sections, tissue cryosections and in cell preparations fixed on slides.

Features - All reagents in dropper bottles prediluted and ready to use - All buffers and enzymes necessary for staining included - Can be used on tissue from any species - Streptavidin-Biotin detection system	Ordering Information
	Cat. No. X2044K1 (pdf) X2044K2 (pdf)	Size 30 Slides 60 Slides
	For larger quantity pricing, please contact Customer Service

Legend:
ApoMark™ kit (X2044K) using paraffin fixed human tonsil tissue, 10 μm sections (1000X). A] Section processed and counter-stained with methyl green according to the ApoMark™ manual. B] Counter-stain step was eliminated to more clearly illustrate the level of positive staining in the germinal centers of tonsil tissue. C] Section treated with DNase I in order to generate a positive control slide. Note all nuclei stain positive. The use of DNase I generates free 3’-OH groups on cellular DNA, these free 3’-OH groups are then labeled with biotin-nucleotide by the TdT in the ApoMark™ kit. D] Negative control, the TdT enzyme step was eliminated thereby generating a negative slide.

Protocol Summary: Not to be used in place of detailed protocol. See kit manual for complete protocol.

Steps	Time (min)
1. Rehydrate samples	25
2. Permeabilize samples using Proteinase K solution provided	20
3. Rinse with TBS	3
4. Inactivate endogenous peroxidases. Incubate with 3% hydrogen peroxide	5
5. Equilibrate samples with equilibration buffer provided	30
6. Add TdT Enzyme to TdT Labeling Reaction Mix and Incubate	90
7. Terminate Reaction with Stop Buffer Provided	5
8. Block Samples with Block Buffer Provided	10
9. Dilute 25x Conjugate provided to 1x and add to each slide	30
10. Rinse with TBS	5
11. Prepare DAB Solution Provided and Add to all samples	15
12. Rinse with dH2O	5
13. Counterstain with Methyl Green Provided	1-3
14. Dehydrate with 100% Methanol – 100% Xylene & mount with coverslip	15