Human NFKB2 (NFkB p100/NFKB2) knockout HCT116 cell line
- Advanced Validation
- What is this?
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Human NFKB2 (NFkB p100/NFKB2) knockout HCT116 cell line available to order. Recommended control: Human wild-type HCT116 cell line (ab255451).
View Alternative Names
CVID10, DNA-binding factor KBF2, H2TF1, LYT 10, Lymphocyte translocation chromosome 10 protein, NFKB p52/p100 subunit, Nuclear factor Kappa B subunit 2, Nuclear factor of kappa light polypeptide gene enhancer in B cells 2 (p49/p100), Nuclear factor of kappa light polypeptide gene enhancer in B-cells 2, Oncogene Lyt-10, Transcription factor NFKB2
- WB
Lab
Western blot - Human NFKB2 (NFkB p100/NFKB2) knockout HCT116 cell line (AB266883)
Lanes 1- 2 : Merged signal (red and green). Green - ab175192 observed at 97 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab175192 was shown to react with NFkB p100/NFKB2 in wild-type HCT116 cells in western blot. The band observed in CRISPR/Cas9 edited cell line ab266883 (CRISPR/Cas9 edited cell lysate ab257245) lane below 97kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HCT116 and NFKB2 CRISPR/Cas9 edited HCT117 cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab175192 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° at a 1 in 10000 dilution and a 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 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686-66] (<a href='/en-us/products/primary-antibodies/nfkb-p100-nfkb2-antibody-epr4686-66-ab175192'>ab175192</a>) at 1/1000 dilution
Lane 1:
Wild-type HCT116 cell lysate at 20 µg
Lane 2:
Western blot - Human NFKB2 (NFkB p100/NFKB2) knockout HCT116 cell line (ab266883)
Lane 2:
NFKB2 CRISPR/Cas9 edited HCT116 cell lysate at 20 µg
Predicted band size: 97 kDa
Observed band size: 120 kDa
false
- WB
Lab
Western blot - Human NFKB2 (NFkB p100/NFKB2) knockout HCT116 cell line (AB266883)
Lanes 1- 2 : Merged signal (red and green). Green - ab109440 observed at 120 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab109440 was shown to react with NFkB p100/NFKB2 in wild-type HCT116 cells in western blot. The band observed in CRISPR/Cas9 edited cell line ab266883 (CRISPR/Cas9 edited cell lysate ab257245) lane below 97kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type HCT116 and NFKB2 CRISPR/Cas9 edited HCT116 cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab109440 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° at a 1 in 1000 dilution and a 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 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-NFkB p100/NFKB2 antibody [EPR4686] (<a href='/en-us/products/primary-antibodies/nfkb-p100-nfkb2-antibody-epr4686-ab109440'>ab109440</a>) at 1/1000 dilution
Lane 1:
Wild-type HCT116 cell lysate at 20 µg
Lane 2:
NFKB2 CRISPR/Cas9 edited HCT116 cell lysate at 20 µg
Lane 2:
Western blot - Human NFKB2 (NFkB p100/NFKB2) knockout HCT116 cell line (ab266883)
Predicted band size: 97 kDa
Observed band size: 120 kDa
false
- Cell Culture
Lab
Cell Culture - Human NFKB2 (NFkB p100/NFKB2) knockout HCT116 cell line (AB266883)
Representative images NFKB2 knockout HCT116 cells, low and high confluency examples (top left and right respectively) and wild-type HCT116 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 M5000 microscope.
- Sanger seq
Unknown
Sanger Sequencing - Human NFKB2 (NFkB p100/NFKB2) knockout HCT116 cell line (AB266883)
Homozygous : 5 bp deletion in exon8
Reactivity data
Product details
Recommended control: Human wild-type HCT116 cell line (ab255451). 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.
We will provide viable cells that proliferate on revival.
Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.
This product is subject to limited use licenses from The Broad Institute and ERS Genomics Limited, and is developed with patented technology. For full details of the limited use licenses and relevant 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
Zygosity
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
McCoY5a + 10% FBS
Cryopreservation medium
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Product protocols
- Visit the General protocols
- Visit the Troubleshooting
Target data
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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