Product Overview
Parental Cell Line
HCT116
Mutation Description
Knockout achieved by using CRISPR/Cas9, homozygous: 1 bp insertion in exon 4.
Knockout Validation
Sanger Sequencing, Western Blot (WB)
Cell Line Properties
Target Information
Gene Function
Tubulin is the major constituent of microtubules. It binds two moles of GTP, one at an exchangeable site on the beta chain and one at a non-exchangeable site on the alpha-chain. TUBB3 plays a critical role in proper axon guidance and mantainance.
Sequence Similarities
Belongs to the tubulin family.
Post-translational Modifications
Some glutamate residues at the C-terminus are polyglutamylated. This modification occurs exclusively on glutamate residues and results in polyglutamate chains on the gamma-carboxyl group. Also monoglycylated but not polyglycylated due to the absence of functional TTLL10 in human. Monoglycylation is mainly limited to tubulin incorporated into axonemes (cilia and flagella) whereas glutamylation is prevalent in neuronal cells, centrioles, axonemes, and the mitotic spindle. Both modifications can coexist on the same protein on adjacent residues, and lowering glycylation levels increases polyglutamylation, and reciprocally. The precise function of such modifications is still unclear but they regulate the assembly and dynamics of axonemal microtubules.
Storage & Handling
Cryopreservation Cell Medium
Cell freezing medium - DMSO serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Culture Medium
McCoY5a + 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.
