BCL6 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 14 bp deletion in exon 2 and Insertion of the selection cassette in exon 2.
Images
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 14 bp deletion in exon 2 and Insertion of the selection cassette in exon 2
Alternative names
B cell CLL/lymphoma 6, B-cell lymphoma 5 protein, B-cell lymphoma 6 protein, BCL-5, BCL6A, BCL6_HUMAN, LAZ 3, LAZ 3 protein, Lymphoma Associated Zinc Finger Gene On Chromosome 3 (LAZ3), Lymphoma associated zinc finger gene on chromosome 3, Protein LAZ-3, ZBTB 27, ZNF 51, Zinc Finger Protein 51 (ZNF51), Zinc finger and BTB domain-containing protein 27, Zinc finger and BTB domain-containing protein 27 (ZBTB27), Zinc finger protein 51, cys-his2 zinc finger transcription factor, zinc finger transcription factor BCL6S
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BCL6 KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 14 bp deletion in exon 2 and Insertion of the selection cassette in exon 2.
Key facts
- Cell type
- HeLa
- Species or organism
- Human
- Tissue
- Cervix
- Form
- Liquid
- Knockout validation
- Sanger Sequencing, Western blot
- Mutation description
- Knockout achieved by using CRISPR/Cas9, 14 bp deletion in exon 2 and Insertion of the selection cassette in exon 2
- Disease
- Adenocarcinoma
- Concentration
Properties
- Gene name
- BCL6
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing, Western blot
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 2 US: 2
- Adherent/suspension
- Adherent
- Gender
- Female
Handling procedures
- 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 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.- Subculture guidelines
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 2x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
- Culture medium
- DMEM (High Glucose) + 10% FBS
- Cryopreservation medium
- Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
The Bcl6 protein also known as B-cell lymphoma 6 is a transcriptional repressor important for the germinal center formation in B-cells. It has a molecular weight of approximately 95 kDa. Bcl6 is expressed mainly in B-cells T-cells and certain subsets of dendritic cells. It functions by binding to target DNA sequences where it represses the expression of specific genes leading to controlled cell proliferation differentiation and apoptosis. Researchers often study Bcl6 through the use of monoclonal antibodies particularly in immunohistochemistry (IHC).
- Biological function summary
Bcl6 plays essential roles in immune response regulation and lymphocyte development. In germinal centers Bcl6 influences the process of somatic hypermutation and class switch recombination in B-cells which is a part of generating antibody diversity. It operates as a central player in a multiprotein complex that includes corepressors such as SMRT and NCoR which facilitate the repression of its target genes. This regulatory action helps maintain the precise balance of immune cell functions.
- Pathways
Bcl6 integrates into the B-cell receptor (BCR) signaling and the JAK-STAT pathway. It interacts with molecules like STAT1 and STAT3 positively influencing the survival and proliferation of B-cells. In these pathways Bcl6's activity modulates the transcriptional landscape that determines cell fate decisions affecting cellular processes such as proliferation and survival in response to external cues.
- Associated diseases and disorders
Bcl6 is relevant to B cell lymphomas specifically diffuse large B-cell lymphoma (DLBCL) where it is often overexpressed contributing to malignancy by preventing cell death. This pathology frequently involves chromosomal translocations that juxtapose the Bcl6 gene with immunoglobulin genes. The relationship between Bcl6 and other proteins like Bcl2 is noteworthy in this context as Bcl2 inhibits apoptosis further contributing to cancer progression. Mutations and dysregulated expression of Bcl6 have also shown links to autoimmune diseases where its role in cell differentiation and survival may lead to aberrant immune responses.
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4 product images
Western blot - Human BCL6 knockout HeLa cell line (ab265410)
Lanes 1-4: Merged signal (red and green). Green Anti-Bcl6 antibody [EP529Y] ab33901 observed at 78 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 37 kDa.Anti-Bcl6 antibody [EP529Y] ab33901 Anti-Bcl6 antibody [EP529Y] was shown to react with BCL6 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265410 (knockout cell lysate Human BCL6 knockout HeLa cell lysate ab257178) was used. Wild-type and Bcl6 knockout samples were subjected to SDS-PAGE. Anti-Bcl6 antibody [EP529Y] ab33901 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4° at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-Bcl6 antibody [EP529Y] (Anti-Bcl6 antibody [EP529Y] ab33901) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: BCL6 knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human BCL6 knockout HeLa cell line (ab265410)
Lane 3: Ramos cell lysate at 20 µg
Lane 4: SH-SY5Y cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 79 kDa
Observed band size: 78 kDa
Cell Culture - Human BCL6 knockout HeLa cell line (ab265410)
Representative images of BCL6 knockout HeLa cells, low and high confluency examples (top left and right respectively) and wild-type HeLa cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS XL Core microscope.
Sanger Sequencing - Human BCL6 knockout HeLa cell line (ab265410)
Allele-2: Insertion of the selection cassette in exon 2.
Sanger Sequencing - Human BCL6 knockout HeLa cell line (ab265410)
Allele-1: 14 bp deletion in exon 2.
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