Key Specifications Table
| Key Applications | Purity |
|---|---|
| Cult | The active component mLIF has been shown to be >95% pure by SDS-PAGE. ESGRO supplement is supplied 0.22 micron filtered, and tested negative in mycoplasma tests. Endotoxin level is less than 0.1 ng per µg of LIF. |
| Description | |
|---|---|
| Catalogue Number | ESG1120 |
| Brand Family | Chemicon® |
| Trade Name |
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| Description | ESGRO®-2i Supplement Kit (1000x) |
| Alternate Names |
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| Background Information | Two stages of mouse pluripotency have been stabilized to date through in vitro culture. ‘Naive’ represents the earliest stage, which can be enriched for a purer population, termed ‘ground state’. Naive most closely approximates embryonic epiblast in the preimplantation blastocyst. The primed state most closely approximates in vivo immediate postimplantation epiblast. There are clear differences between these stages in culture, expression, epigenetics (including appropriate X-inactivation) and developmental competence. The fact that naive mESCs can generate whole animals through tetraploid complementation while primed mESCs cannot, is a particularly compelling reason for heightened expectations from naive hESCs. Mouse Leukemia Inhibitory Factor (LIF) is a critical component in mouse ES cell culture media where it promotes the self-renewal and maintains the pluripotency of mouse embryonic stem (ES) cells. Recent studies have determined that inhibitors that block MAPK/Erk pathway, in combination with the glycogen synthase kinase 3 (GSK3) inhibitor, protect mouse ES cells from differentiation-inducing signals, allowing for self-renewal even in the absence of LIF. Mouse ES cells cultured in these conditions maintain their LIF responsiveness and studies have shown that adding LIF to the 2i culture conditions led better colony propagation and enhanced the general health of ES colonies. Supports a naive/ground pluripotent state. |
| Product Information | |
|---|---|
| Components |
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| Presentation | Both supplements were formulated at 1000x concentration in solution. Mouse LIF is prepared in phosphate buffered saline with 1% w/v bovine serum albumin BSA as a carrier for stability. MEK/GSK3 inhibitor supplement is prepared in DMSO. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | The ESGRO®-2i Supplement Kit (1000x) contains LIF a selective GSK3β & Mek 1/2 inhibitors to enhance viability of mouse embryonic stem & iPS cells in the absence of serum & feeder cells. Supports a naive/ground pluripotent state. |
| Key Applications |
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| Application Notes | Suggested Protocol for Medium Preparation: 1. Thaw MEK/GSK3 inhibitor supplement and mLIF at room temperature. 2. Each vial of supplement is sufficient to make 500 mL of complete medium for serum-free or serum dependent mouse ES culture. 3. Aseptically transfer the contents of the vials containing the supplements at 1000x dilution to the pre-warmed bottle of serum free basal medium such as ESGRO Complete Basal Medium (Cat. No. SF002-500) or serum containing medium and mix. 4. The complete medium can be filtered through 0.22 µm filter (Cat. No. SCGVU05RE). 5. After supplements are added to the medium, store in the dark at 4°C for up to ten days. |
| Biological Information | |
|---|---|
| Purity | The active component mLIF has been shown to be >95% pure by SDS-PAGE. ESGRO supplement is supplied 0.22 micron filtered, and tested negative in mycoplasma tests. Endotoxin level is less than 0.1 ng per µg of LIF. |
| Specific Activity | ESGRO Supplement is assessed both on mouse ES cells and on murine M1 myeloid leukemic cells. A standard of 50 Units is defined as the concentration of ESGRO supplement in 1.0 mL of tissue culture medium that induces the differentiation of 50% of M1 colonies (Metcalf, 1988).<br /><br />Embryonic Stem Cell Assay: Differentiation inhibition at 1000 units/mL<br />Murine myeloid leukemic, M1 Assay: Specific Activity is greater than or equal to 10<sup>8</sup> units/mg |
| Entrez Gene Number |
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| Gene Symbol |
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| UniProt Number |
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| Media Form | Liquid |
| Stem Cell Type |
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| Product Usage Statements | |
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| Usage Statement |
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| Storage and Shipping Information | |
|---|---|
| Storage Conditions | Store at -20°C. Upon thaw, ESGRO Supplement should be stored at 2-8°C. Do not refreeze ESGRO Supplement (part no. CS210511). Refer to label for expiration date. |
| Packaging Information | |
|---|---|
| Material Size | 1 kit per 5 L medium |
| Material Package | Provided in ready to use aliquots sufficient for supplementing 10 x 500 mL mouse ES cell culture medium for a total of 5 liters complete medium. |