人NFKB2 (NFkB p100/NFKB2) knockout HCT116 cell line
Human NFKB2 (NFkB p100/NFKB2) knockout HCT116 cell line
- Advanced Validation
- 了解详情
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NFKB2 KO cell line available to order. KO validated. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 5 bp deletion in exon 8. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
查看别名
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 - 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='/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
- 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='/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
- Sanger seq
Unknown
Sanger Sequencing - Human NFKB2 (NFkB p100/NFKB2) knockout HCT116 cell line (AB266883)
Homozygous : 5 bp deletion in exon8
- 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.
反应性数据
产品详情
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.
规格
性能和储存信息
基因名称
基因编辑类型
基因编辑方法
敲除验证
合子性
运输条件
推荐的短期储存条件
推荐的长期储存条件
处理步骤
初始处理指南
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.
传代培养指南
- 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.
培养基
McCoY5a + 10% FBS
低温储藏试剂
Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.
质量控制
STR 分析
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
细胞培养
生物安全等级
EU: 1 US: 1
贴壁/悬浮
Adherent
性别
Male
搭配使用产品
Primary Antibodies
AB175192
Anti-NFkB p100/NFKB2 antibody [EPR4686-66]
RabMAb|Recombinant|KO Validated
![Immunocytochemistry/ Immunofluorescence - Anti-NFkB p100/NFKB2 antibody [EPR4686-66] (AB175192)](https://content.abcam.com/products/images/nfkb-p100-nfkb2-antibody-epr4686-66-ab175192--immunocytochemistry-immunofluorescence-img123054.jpg)
- 5
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Cell Lines & Lysates
AB264673
Human PRKACB (PKA beta catalytic subunit) knockout HeLa cell line
Advanced Validation
- 0
- 0
Abcam 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