Human BCL6 knockout HeLa cell line
- Advanced Validation
- What is this?
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(0 Publication)
- WB
Lab
Western blot - Human BCL6 knockout HeLa cell line (AB265410)
Lanes 1-4 : Merged signal (red and green). Green ab33901 observed at 78 kDa. Red - loading control ab8245 observed at 37 kDa.
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 ab257178) was used. Wild-type and Bcl6 knockout samples were subjected to SDS-PAGE. ab33901 and 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 (ab216773) and 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] (<a href='/en-us/products/primary-antibodies/bcl6-antibody-ep529y-ab33901'>ab33901</a>) 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
Predicted band size: 79 kDa
Observed band size: 78 kDa
false
- Cell Culture
Unknown
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 seq
Unknown
Sanger Sequencing - Human BCL6 knockout HeLa cell line (AB265410)
Allele-1 : 14 bp deletion in exon 2.
- Sanger seq
Unknown
Sanger Sequencing - Human BCL6 knockout HeLa cell line (AB265410)
Allele-2 : Insertion of the selection cassette in exon 2.
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Cell Lines & Lysates
AB255358
Human TUBB3 (beta III Tubulin) knockout HeLa cell line
Advanced Validation
- 0
- 0
Reactivity data
Product details
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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
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.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com