人IL-6 knockout A549 cell line
Human IL-6 knockout A549 cell line
- Advanced Validation
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IL6 KO cell line available to order. KO validated by Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 97 bp deletion in exon 3. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
查看别名
B cell differentiation factor, B-cell stimulatory factor 2, BSF-2, CTL differentiation factor, Cytotoxic T cell differentiation factor, HSF, Hepatocyte stimulating factor, Hepatocyte stimulatory factor, Hybridoma growth factor, Hybridoma growth factor Interferon beta-2, Hybridoma plasmacytoma growth factor, IFN-beta-2, IFNB2, IL6_HUMAN, Interferon beta-2, Interleukin 6 (interferon beta 2), Interleukin BSF 2, Interleukin-6
- WB
Lab
Western blot - Human IL-6 knockout A549 cell line (AB273751)
Lanes 1 - 4 : Merged signal (red and green). Green - ab233706 observed at 25 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.
ab233706 was shown to react with IL-6 in wild-type A549 cells in western blot with loss of signal observed in IL-6 knockout cell line ab273751 (knockout cell lysate ab275501). Wild-type and IL-6 knockout A549 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab233706 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4° at a 1 in 1000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-IL-6 antibody [EPR21711] (<a href='/products/primary-antibodies/il-6-antibody-epr21711-ab233706'>ab233706</a>) at 1/1000 dilution
Lane 1:
Wild-type A549 Brefeldin A (<a href='/products/biochemicals/brefeldin-a-inhibitor-of-adp-ribosylation-factor-ab120299'>ab120299</a>)-treated (5ug/ml, 4h) cell lysate at 30 µg
Lane 2:
Wild-type A549 IL-1β (<a href='/products/proteins-peptides/recombinant-human-il-1-beta-protein-active-ab259387'>ab259387</a>) (20 ng/ml, 24h) and Brefeldin A (<a href='/products/biochemicals/brefeldin-a-inhibitor-of-adp-ribosylation-factor-ab120299'>ab120299</a>)-treated (5 ug/ml for the last 4h) cell lysate at 30 µg
Lane 2:
Western blot - Human IL-6 knockout A549 cell line (ab273751)
Lane 3:
IL-6 knockout A549 Brefeldin A (<a href='/products/biochemicals/brefeldin-a-inhibitor-of-adp-ribosylation-factor-ab120299'>ab120299</a>)-treated (5ug/ml, 4h) cell lysate at 30 µg
Lane 4:
IL-6 knockout A549 IL-1β (<a href='/products/proteins-peptides/recombinant-human-il-1-beta-protein-active-ab259387'>ab259387</a>) (20 ng/ml, 24h) and Brefeldin A (<a href='/products/biochemicals/brefeldin-a-inhibitor-of-adp-ribosylation-factor-ab120299'>ab120299</a>)-treated (5 ug/ml for the last 4h) cell lysate at 30 µg
Predicted band size: 23 kDa
Observed band size: 25 kDa,35 kDa,40 kDa
false
- WB
Lab
Western blot - Human IL-6 knockout A549 cell line (AB273751)
Lanes 1 - 4 : Merged signal (red and green). Green - ab214429 observed at 25 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.
ab214429 was shown to react with IL-6 in wild-type A549 cells in western blot with loss of signal observed in IL-6 knockout cell line ab273751 (knockout cell lysate ab275501). Wild-type and IL-6 knockout A549 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab214429 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4° at a 1 in 1000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-IL-6 antibody [EPR20653] (<a href='/products/primary-antibodies/il-6-antibody-epr20653-ab214429'>ab214429</a>) at 1/1000 dilution
Lane 1:
Wild-type A549 Brefeldin A (<a href='/products/biochemicals/brefeldin-a-inhibitor-of-adp-ribosylation-factor-ab120299'>ab120299</a>)-treated (5ug/ml, 4h) cell lysate at 30 µg
Lane 2:
Wild-type A549 IL-1β (<a href='/products/proteins-peptides/recombinant-human-il-1-beta-protein-active-ab259387'>ab259387</a>) (20 ng/ml, 24h) and Brefeldin A (<a href='/products/biochemicals/brefeldin-a-inhibitor-of-adp-ribosylation-factor-ab120299'>ab120299</a>)-treated (5 ug/ml for the last 4h) cell lysate at 30 µg
Lane 2:
Western blot - Human IL-6 knockout A549 cell line (ab273751)
Lane 3:
IL-6 knockout A549 Brefeldin A (<a href='/products/biochemicals/brefeldin-a-inhibitor-of-adp-ribosylation-factor-ab120299'>ab120299</a>)-treated (5ug/ml, 4h) cell lysate at 30 µg
Lane 4:
IL-6 knockout A549 IL-1β (<a href='/products/proteins-peptides/recombinant-human-il-1-beta-protein-active-ab259387'>ab259387</a>) (20 ng/ml, 24h) and Brefeldin A (<a href='/products/biochemicals/brefeldin-a-inhibitor-of-adp-ribosylation-factor-ab120299'>ab120299</a>)-treated (5 ug/ml for the last 4h) cell lysate at 30 µg
Predicted band size: 23 kDa
Observed band size: 25 kDa,35 kDa
false
- sELISA
Lab
Sandwich ELISA - Human IL-6 knockout A549 cell line (AB273751)
Human IL-6 concentration was interpolated from the IL-6 standard curve. Supernatants from cell culture samples were serially diluted and assessed by the Human IL-6 ELISA kit (ab178013). Wild-type and IL-6 knockout A549 cells (ab273751) were assessed in duplicate (n=2). Cells were either treated with 20 ng/mL active recombinant human IL-1 beta protein (ab259387) for 24 h to induce expression of IL-6 or not treated with IL-1 beta. Data are represented as the mean and error bars represent standard deviation.
- Sanger seq
Lab
Sanger Sequencing - Human IL-6 knockout A549 cell line (AB273751)
Allele-1 : 97 bp deletion in exon 3.
反应性数据
产品详情
We will provide viable cells that proliferate on revival.
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.
- Do not allow the cell density to exceed 7x104 cells/cm2.
培养基
F-12K + 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.
补充信息
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
IL-6 influences immune regulation and acts as part of the acute phase response. It stimulates the production of acute-phase proteins and supports the differentiation of B cells into antibody-producing cells. IL-6 is not known to be part of a larger complex acting primarily as a single entity in signal transduction. Moreover IL-6 impacts the metabolism of iron and bone homeostasis showing its multifunctional nature.
Pathways
IL-6 forms an integral part of several signaling routes particularly the JAK-STAT pathway. In this context IL-6 interacts with signal transducer proteins like STAT3 to transmit signals from the cell surface to the nucleus affecting gene expression. Another important pathway is the MAPK pathway through which IL-6 influences cell proliferation and survival. These interactions reflect IL-6's diverse effects in cellular processes.
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
AB259341
Anti-IL-6 antibody [EPR23819-11]
20ul selling size|RabMAb|Recombinant
![Immunoprecipitation - Anti-IL-6 antibody [EPR23819-11] (AB259341)](https://content.abcam.com/products/images/il-6-antibody-epr23819-11-ab259341--immunoprecipitation-img164278.jpg)
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