Product Overview
Parental Cell Line
RAW 264.7
Mutation Description
Knockout achieved by using CRISPR/Cas9, homozygous: 52 bp deletion-3 bp insertion in Exon 1
Knockout Validation
Sanger Sequencing, Western Blot (WB)
Cell Line Properties
Morphology
Macrophage-like
Target Information
Gene Function
Important regulator of cell cycle progression. Involved in G1 arrest. Potent inhibitor of cyclin E- and cyclin A-CDK2 complexes. Forms a complex with cyclin type D-CDK4 complexes and is involved in the assembly, stability, and modulation of CCND1-CDK4 complex activation. Acts either as an inhibitor or an activator of cyclin type D-CDK4 complexes depending on its phosphorylation state and/or stoichometry.
Sequence Similarities
Belongs to the CDI family.
Post-translational Modifications
Phosphorylated; phosphorylation occurs on serine, threonine and tyrosine residues. Phosphorylation on Ser-10 is the major site of phosphorylation in resting cells, takes place at the G(0)-G(1) phase and leads to protein stability. Phosphorylation on other sites is greatly enhanced by mitogens, growth factors, cMYC and in certain cancer cell lines. The phosphorylated form found in the cytoplasm is inactivate. Phosphorylation on Thr-198 is required for interaction with 14-3-3 proteins. Phosphorylation on Thr-187, by CDK2 leads to protein ubiquitination and proteasomal degradation. Tyrosine phosphorylation promotes this process. Phosphorylation by PKB/AKT1 can be suppressed by LY294002, an inhibitor of the catalytic subunit of PI3K. Phosphorylation on Tyr-88 and Tyr-89 has no effect on binding CDK2, but is required for binding CDK4. Dephosphorylated on tyrosine residues by G-CSF.Ubiquitinated; in the cytoplasm by the KPC complex (composed of RNF123/KPC1 and UBAC1/KPC2) and, in the nucleus, by SCF(SKP2). The latter requires prior phosphorylation on Thr-187. Ubiquitinated; by a TRIM21-containing SCF(SKP2)-like complex; leads to its degradation.Subject to degradation in the lysosome. Interaction with SNX6 promotes lysosomal degradation.
Storage & Handling
Cryopreservation Cell Medium
Cell freezing medium - DMSO serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Culture Medium
DMEM + 10% FBS
Initial Handling Guidelines
Lyophilizate may be stored at 4 °C. After reconstitution, store at -20 °C for short-term storage or -80 °C for long-term storage.
Storage Instructions
Store at -80 °C. Avoid freeze / thaw cycles.
