Amersham™ AlkPhos direct hybridization buffer
AlkPhos Direct Labeling and Detection Systems are based on the rapid, direct labeling of DNA or RNA probes with thermostable alkaline phosphatase. CDP-Star or ECF detection can be used.
Amersham™ AlkPhos direct hybridization buffer
AlkPhos Direct Labeling and Detection Systems are based on the rapid, direct labeling of DNA or RNA probes with thermostable alkaline phosphatase. CDP-Star or ECF detection can be used.
Overview
AlkPhos Direct Labeling and Detection Systems are based on the rapid, direct labeling of DNA or RNA probes with thermostable alkaline phosphatase. CDP-Star or ECF detection can be used.
- Fast and easy to use system suitable for the majority of routine blotting applications, using either DNA, RNA or oligonucleotide probes.
- Alk Phos Direct system has less components than other non-radioactive systems, making it simpler to use as less components need to be removed during the stripping procedure.
- Probe removal is more effective with less damage to the membrane.
- Optimized protocol has been developed capable of labelling probes down to 17 base pairs in length.
- 3–4 hours of time savings compared with conventional, indirect methods are achievable as a result of the rapid and simple labelling reaction.
Denatured or single-stranded DNA or RNA probe is mixed with the labelling buffer and alkaline phosphatase. The probe is hybridized to the blot using the specially formulated AlkPhos Direct Hybridization buffer, which stabilizes the activity of the enzyme. After a 15-minute pre-hybridization step, hybridization with probe is performed overnight at the required temperature. For higher target amounts, a 2–4 hour hybridization may be sufficient.
