Product Size
1 x 106 cells/vial, 1 mL
Product Overview
Mutation Description
Knockout achieved by using CRISPR/Cas9, 139 bp insertion in exon 2 and 2 bp deletion in exon 2
Knockout Validation
Sanger Sequencing, Western Blot (WB)
Cell Line Properties
Adherent / Suspension
Adherent
Target Information
Gene Function
Ser/Thr-kinase component of cyclin D-CDK4 (DC) complexes that phosphorylate and inhibit members of the retinoblastoma (RB) protein family including RB1 and regulate the cell-cycle during G(1)/S transition. Phosphorylation of RB1 allows dissociation of the transcription factor E2F from the RB/E2F complexes and the subsequent transcription of E2F target genes which are responsible for the progression through the G(1) phase. Hypophosphorylates RB1 in early G(1) phase. Cyclin D-CDK4 complexes are major integrators of various mitogenenic and antimitogenic signals. Also phosphorylates SMAD3 in a cell-cycle-dependent manner and represses its transcriptional activity. Component of the ternary complex, cyclin D/CDK4/CDKN1B, required for nuclear translocation and activity of the cyclin D-CDK4 complex.
Sequence Similarities
Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. CDC2/CDKX subfamily.Contains 1 protein kinase domain.
Post-translational Modifications
Phosphorylation at Thr-172 is required for enzymatic activity. Phosphorylated, in vitro, at this site by CCNH-CDK7, but, in vivo, appears to be phosphorylated by a proline-directed kinase. In the cyclin D-CDK4-CDKN1B complex, this phosphorylation and consequent CDK4 enzyme activity, is dependent on the tyrosine phosphorylation state of CDKN1B. Thus, in proliferating cells, CDK4 within the complex is phosphorylated on Thr-172 in the T-loop. In resting cells, phosphorylation on Thr-172 is prevented by the non-tyrosine-phosphorylated form of CDKN1B.
Storage & Handling
Recommended Control
Human wild-type HeLa cell line.
Cryopreservation Cell Medium
Cell freezing medium - DMSO serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Culture Medium
DMEM (High Glucose) + 10% FBS
Initial Handling Guidelines
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80 °C. Storage at -80 °C may result in loss of viability.
Storage Instructions
Shipped on dry ice. Store in liquid nitrogen.
Storage Buffer
Constituents: 8.7% Dimethylsulfoxide, 2% Cellulose, methyl ether.
