Cre Recombinase (Cre, Cre (Cyclization Recombination) Enzyme)

The Cre enzyme, a member of a large family of recombinases, recognizes loxP which is a sequence motif of 34bp from the PI bacteriophage. If a DNA seqment is flanked by two loxP sites in the same orientation, Cre excises that segment leaving a single loxP site in the remaining DNA molecule. The excised DNA segment is subsequently degraded. Cre and lox comprise the Cre/lox system which was first developed in the late 1980’s to artificially manipulate gene expression. As an example, mice with the Cre protein expressed in a specific cell type are bred with mice that contain a target gene surrounded by loxP sites. When the mice are bred, the cells carrying Cre will cause those cells to lose the target gene. If the Cre gene is, for example, bound to a promoter that only allows Cre production in neuronal cells, the target gene will be deleted only in those cells. There are numerous animal, plant and bacterial stocks that already contain the Cre gene driven by ubiquitous, tissue-specific or inducible promoters and provide a quick method for breeding experiments. The Cre/lox system has the advantage of working in almost any type of cell. This has led to its application in many experiments, for example in selectively labeling neuronal cells in the brain thereby differentiating them from other types of surrounding cells.

Applications:
Suitable for use in Western Blot, Immunoprecipitation, Immunofluorescence/Immunocytochemistry and Immunohistochemistry. Other applications not tested.

Recommended Dilutions:
Western Blot: 1:1000-1:4000
Immunoprecipitation: 1:50-1:200
Immunofluorescence (IC): 1:10-1:500
Immunohistochemistry (Paraffin): 1:1000-1:5000
Immunohistochemistry (Frozen): 1:1000-1:5000
Optimal dilutions to be determined by the researcher.

Positive Controls:
Recombinant CRE, Cells stably transfected with CRE, Tissue from a CRE transgenic organism

Storage and Stability:
May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 12 months after receipt. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.