AB273751

Human IL-6 knockout A549 cell line

Advanced Validation
詳細を見る

Be the first to review this product! Submit a review

(0 Publication)

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 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.

別名を表示する

IFN-beta-2, CTL differentiation factor, CDF, B-cell stimulatory factor 2, BSF-2, IL-6, Interleukin-6, Hybridoma growth factor, IFNB2, Interferon beta-2, IL6

4 Images

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

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:

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:

Predicted band size: 23 kDa

Observed band size: 25 kDa,35 kDa

false

Sandwich ELISA - Human IL-6 knockout A549 cell line (AB273751)

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 Sequencing - Human IL-6 knockout A549 cell line (AB273751)

Sanger seq

Lab

Sanger Sequencing - Human IL-6 knockout A549 cell line (AB273751)

Allele-1 : 97 bp deletion in exon 3.

Key facts

細胞タイプ

A549

生物種

Human

組織

Lung

製品の状態

Liquid

form

ノックアウト検証方法

Sanger Sequencing,Western blot

ノックアウト変異

Knockout achieved by using CRISPR/Cas9, Homozygous: 97 bp deletion in exon 3

疾病

Carcinoma

style

left-column

style

left-column

Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"" }, "sELISA": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"" } } }

style

left-column

製品の詳細

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.

style

left-column

製品内容

{ "values": { "2x1000000Cellsvial": { "sellingSize": "2 x 1000000 Cells/vial", "publicAssetCode":"ab273751-2x1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab273751 Human IL-6 knockout A549 cell line", "number":"AB273751-CMP01" }, { "size":"1 x 1000000 Cells/vial", "name":"ab275463 Human wild-type A549 cell line", "number":"AB273751-CMP02" } ] }, "1000000Cellsvial": { "sellingSize": "1000000 Cells/vial", "publicAssetCode":"ab273751-1000000Cells_vial", "assetComponentDetails": [ { "size":"1 x 1000000 Cells/vial", "name":"ab273751 Human IL-6 knockout A549 cell line", "number":"AB273751-CMP01", "productcode":"" } ] } } }

style

left-column

出荷温度及び保存条件

遺伝子名

IL6

遺伝子編集のタイプ

Knockout

遺伝子編集の方法

CRISPR technology

ノックアウト検証方法

Sanger Sequencing, Western blot

接合型

Homozygous

出荷温度

Dry Ice

短期保存温度

-196°C

長期保存温度

-196°C

style

left-column

補足情報

This supplementary information is collated from multiple sources and compiled automatically.

Interleukin-6 (IL-6) a cytokine also known as IFN-beta2 plays a significant role in immune response and inflammation. The IL-6 protein has a molecular weight of approximately 20-26 kDa. Expression of IL-6 occurs in various cell types including T cells macrophages and fibroblasts. Researchers often measure IL-6 levels in biological samples using IL-6 ELISA kits an essential tool for studying this protein's function and presence in experimental and clinical settings.

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.

IL-6's association with rheumatoid arthritis and multiple myeloma emphasizes its role in chronic inflammation and cancer. In rheumatoid arthritis IL-6 contributes to inflammation and joint damage often together with TNF-alpha highlighting a potential target for anti-inflammatory therapies. In multiple myeloma IL-6 supports the survival and proliferation of cancerous plasma cells highlighting its importance in cancer progression and possible treatment targets.

style

left-column

style

left-column

Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

style

left-column

Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Male

初回取り扱いガイドライン

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.

継代培養ガイドライン

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.

培養培地

F-12K + 10% FBS

凍結保存培地

Cell Freezing Medium-DMSO Serum free media, contains 8.7% DMSO in MEM supplemented with methyl cellulose.

style

left-column

製品プロトコール

Visit the General protocols
Visit the Troubleshooting

style

left-column

ターゲットの情報

See full target information IL6

style

left-column

style

left-column

style

right-column

Abcam product promise

当社は、高品質な試薬を通じてお客様の研究を力強くサポートすることをお約束いたします。ご使用いただく各段階で、常にお客様をサポートできる体制を整えております。万が一、製品が期待通りに機能しない場合は、「Abcam Product Promise」による当社保証制度に基づき、安心してご利用いただけます。

保証に関する詳細については利用規約をご確認ください。

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

Human IL-6 knockout A549 cell line