ZBTB33 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Insertion of the selection cassette in exon 2.
ZBTB33 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Insertion of the selection cassette in exon 2.
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.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.
Although we aim to provide customers with a homozygous clone, feasibility will be dependent on the biology of the protein. Should only heterozygous edits be achieved, you will be notified of the outcome and be asked to confirm whether the cell line is acceptable. All clones will be accompanied with DNA sequencing data, and the mutation description.
Recommended control: Human wild-type HeLa cell line (ab255448). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.
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.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x10E4 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.
Subculture guidelines:
• All seeding densities should be based on cell counts gained by established methods.
• A guide seeding density of 2x10E4 cells/cm2 is recommended.
• Cells should be passaged when they have achieved 80-90% confluence.
• Do not allow the cell density to exceed 7x10E4 cells/cm2.
We will provide viable cells that proliferate on revival.
This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.
Kaiso also known as ZBTB33 is a transcriptional regulator with a mass of approximately 75 kilodaltons. It is a member of the POZ-zinc finger family of proteins. Researchers identified Kaiso as a dual-specificity transcription factor meaning it can bind to both DNA methylated and non-methylated sequences. It is widely expressed in various tissues including the brain liver heart and testis. This broad expression pattern indicates its potential involvement in multiple cellular processes.
Kaiso functions as a transcriptional repressor influencing gene expression by binding to specific DNA sequences. It operates as part of a protein complex that includes other corepressors like N-CoR and SMRT. These interactions modulate chromatin structure which affects gene accessibility. Kaiso's repression of target genes is essential for maintaining cellular homeostasis. Research shows that Kaiso also interacts with p120-catenin a protein involved in cell adhesion linking its function to cytoskeletal dynamics and cell motility.
Several studies show that Kaiso is part of key signaling cascades such as the Wnt/β-catenin pathway. It interacts with other pathway regulators including TCF/LEF transcription factors and is influenced by β-catenin signaling which plays an important role in embryonic development and cellular proliferation. Another significant pathway involving Kaiso is the Notch signaling pathway where Kaiso's function contributes to cell fate determination and differentiation. Through these pathways its ability to regulate gene expression impacts many biological processes.
Kaiso has been linked to cancer progression and metastasis. Its downregulation or loss of function can lead to tumor growth primarily due to the deregulation of genes involved in cell proliferation and apoptosis. Studies highlight its association with colorectal cancer where it influences the expression of tumor suppressor genes. Additionally Kaiso's interaction with β-catenin connects it to other diseases like colorectal cancer where dysregulation of the Wnt pathway plays a critical role. These connections emphasize the potential impact of Kaiso on disease pathogenesis and its value as a therapeutic target.
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