Stem-Molecule — CHIR99021
CHIR99021 is a selective inhibitor of glycogen synthase kinase 3. CHIR99021 has been used to propagate mouse embryonic stem cells in 2i or 3i culture and is used to derive and propagate extended pluripotent stem (EPS) cell culture for mouse, human and pig. CHIR99021 can also be used for expansion of human naïve pluripotent cells containing the genetic pluripotency reporter OCT4-ΔPE-GFP+.
25 mM in DMSO
25 mM in DMSO
25 mM in DMSO
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Product Literature & Protocol
FAQs & Troubleshooting
- 2i or 3i mouse embryonic stem cell culture
- Media component of extended pluripotent stem (EPS) culture for mouse, human and pig
- Generation of chemically reprogrammed mouse induced pluripotent stem cells
- Derivation of nascent ES cell lines from epiblast/ICM from difficult and non-permissive mouse strains
- Component of naïve human pluripotent stem cell media including t2iLGöY media
This product is manufactured in the USA using chemically-defined components and, therefore, does not contain manufacturing components such as serum or animal-derived raw materials.
>99 % free base. FTIR (Fourier Transform Infrared Spectroscopy) analysis was used to compare peaks in infrared spectrum with those of a known standard to determine trace impurities.
|Molecular Weight||465.34 g/mol|
Stock solutions of CHIR99021 in its concentrated form can be stored at least 6 months at -20°C and at least 12 months at -80°C from the date of manufacture with no loss of activity. For long term storage, avoid repeated freeze-thaw cycles by aliquoting. Multiple freeze-thaw cycles reduce potency and, therefore, aliquoting working stocks is strongly recommended.
A sterile, aqueous, clear and colorless solution.
CHIR99021 is supplied at 25 mM in the excipient DMSO. The reconstituted solution has been filter sterilized by passing through a 0.22 micron PES membrane and tested to be negative for mycoplasma contamination.
Disclaimer & Precautions:
This product is solely for research and development use only and may be subject to conditional use and licensing restrictions. The product shall not be used as an advanced pharmaceutical intermediate (API) or investigational drug or a biologic. This product is not intended to be used as a therapeutic agent or facilitate clinical diagnosis or be used as an in vitro diagnostic (IVD) product.
The Food and Drug Administration (FDA) and Center for Biologics Evaluation and Research (CBER) define an IVD as:
“In vitro diagnostic products are those reagents, instruments, and systems intended for use in the diagnosis of disease or other conditions, including a determination of the state of health, in order to cure, mitigate, treat, or prevent disease or its sequelae. Such products are intended for use in the collection, preparation, and examination of specimens taken from the human body. These products are devices as defined in section 201(h)of the Federal Food, Drug, and Cosmetic Act (the act), and may also be biological products subject to section 351 of the Public Health Service Act. Title 21, Code of Federal Regulations (CFR), section 809.3(a).”
This product shall not be used or formulated in any agricultural, pesticidal, veterinary or animal products, food additives or household chemicals or any other unspecified use. Please consult the Safety Data Sheet for information regarding hazards and safe handling practices. NovoHelix distributes products for basic and translational research use only. NovoHelix will report any unspecified use to respective regulatory authorities for enforcement to ensure safeguarding of our research products from potential abuse.
Notice to purchaser:
The purchase price of this product includes a limited, non-transferable license under U.S. and foreign patents or applications owned by NovoHelix to use this product. No other license under these patents or applications is conveyed expressly or by implication by purchase of this product.
Product Literature & Protocol
- HiFi DNA Assembly Protocol
Recommended Amount of Fragments Used for Assembly
|2–3 Fragment Assembly*||4–6 Fragment Assembly**||Positive Control✝|
|Recommended DNA Molar Ratio||vector:insert = 1:2||vector:insert = 1:1|
|Total Amount of Fragments|
HiFi DNA Assembly Master Mix
|10 μl||10 μl||10 μl|
|Deionized H2O||10-X μl||10-X μl||0|
|Total Volume||20 μl✝✝||20 μl✝✝||20 μl|
* Optimized cloning efficiency is 50–100 ng of vector with 2-fold excess of inserts.
Use 5 times more insert if size is less than 200 bp. Total volume of unpurified PCR fragments in the assembly reaction should not exceed 20%.
**To achieve optimal assembly efficiency, design ≥ 20 bp overlap regions between each fragment with equimolarity (suggested: 0.05 pmol each).
† Control reagents are provided for 5 experiments.
†† If greater numbers of fragments are assembled, increase the volume of the reaction linearly by using additional NovoHelix HiFi DNA Assembly Master Mix. Alternatively, pool the DNA fragments into an equimolar mix first and then re-purify these pooled equimolar fragments over a micro-column and elute with a minimum volume (~10-µl). The eluate may be reapplied to the same micro-column membrane to improve elution of large DNA fragments without increasing the final volume..
Recommended Storage Condition:
This assembly mixture can be stored at -20 °C for at least one year. The enzymes remain active following at least 10 freeze-thaw cycles.
FAQs & Troubleshooting
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