Recombinant Afamin/Wnt3a

Recombinant Wnt3a protein and Afamin protein. The N-terminal PA tagged mouse Wnt3a and the N-terminal Myc tagged human Afamin were co-expressed in CHO-K1 cells.

Note: We recommend the addition of albumin to dilution buffer to improve freeze-thaw stability, not diluted without albumin such as PBS only, if diluted product is stored in freeze. Dilutions prepared by the following protocol have been shown to maintain high Wnt3a activity after a freeze-thaw cycle.
1. Prepare PBS containing 0.04% HSA (human serum albumin) or BSA (bovine serum albumin).
2. Dilute Recombinant Afamin/Wnt3a with 0.04% HSA or BSA/PBS to prepare 2-, 10-, and 50-fold dilutions.
Use the prepared dilutions as soon as possible and avoid repeated freeze-thaw cycles.

Note Regarding Intellectual Property Rights:

When culturing organoids, or stem cells, or other tissues, if you are to use this product (J2-001 Afamin/Wnt3a CM) in combination with another factor or factors (hereinafter “factors”), a third party may have patent rights concerning the use or other application of the factors concerned.  MBL does not offer any non-infringement warranty for J2-001 Afamin/Wnt3a CM when used or otherwise applied in combination with other factors. Therefore, if you intend to use this product in combination with other factors, please check with your organization’s division or advisors responsible for intellectual property rights or with your research agency before using this product with other factors.

Product Type: Cell Culture

Application: Cell Culture

Formulation: 60 ug in 300 uL volume of 20 mM Tris-HCI (pH 7.4), 150 mM NaCI

Research Area: Organoid

Description/Background: Wnt signaling is known to be involved in early development, maintenance and regeneration of stem cells, and in cancer formation. Wnt signaling has also been found to play an important role in the growth and maintenance of these processes. In particular, Wnt3a has been revealed to be an essential niche component for maintaining the proliferation of Lgr5-positive stem cells in intestinal epithelial cells and is used for the production of various digestive organoids such as the small intestine, large intestine, stomach, pancreas and liver. Although Wnt3a has been conventionally used for the culture of gut organoids, it is a fat-soluble protein, so it forms aggregates in serum-free medium and cannot exert its activity sufficiently. In 2016, Mihara et al. found that high Wnt3a activity can be maintained by forming a complex with Wnt3a by Afamin, which is one of the components of serum. In addition, by using Afamin and Wnt3a complex for organoid culture, long-term culture of organoid becomes possible. This new medium will result in optimal success for your organoid experiments.

Storage Temperature: -20°C

Protocols: Cell Culture

Purity: Greater than 90% purity as confirmed on SDS-PAGE

Endotoxin levels: < 1.0 EU/mL by the LAL assay

Regulatory Statement: For Research Use Only. Not for use in diagnostic procedures.