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AB266873

人ADAM17 knockout HCT116 cell line

Human ADAM17 knockout HCT116 cell line

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ADAM17 KO cell line available to order. KO validated. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 5. 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

ADAM17, TACE, CSVP, ADAM 17, Snake venom-like protease, TNF-alpha-converting enzyme, Disintegrin and metalloproteinase domain-containing protein 17, CD156b, TNF-alpha convertase

2 Images
Western blot - Human ADAM17 knockout HCT116 cell line (AB266873)
  • WB

Lab

Western blot - Human ADAM17 knockout HCT116 cell line (AB266873)

Lanes 1 - 2 : Merged signal (red and green). Green - ADAM17 observed at 120 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa. Rabbit anti-ADAM17 antibody was shown to react with TACE in wild-type cells in Western blot with loss of signal observed in ADAM17 knockout cell line ab266873 (ADAM17 knockout cell lysate ab256832). Wild-type and ADAM17 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with rabbit anti-ADAM17 antibody and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C 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 h at room temperature before imaging.

All lanes:

Western blot - Anti-alpha Tubulin antibody [DM1A] - Loading Control (<a href='/products/primary-antibodies/alpha-tubulin-antibody-dm1a-loading-control-ab7291'>ab7291</a>) at 1/1000 dilution

Lane 1:

Wild-type cell lysate

Lane 2:

Western blot - Human ADAM17 knockout HCT116 cell line (ab266873)

Secondary

Lane 1:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/20000 dilution

Lane 2:

Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (<a href='/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-680rd-preadsorbed-ab216776'>ab216776</a>) at 1/20000 dilution

Predicted band size: 93 kDa

Observed band size: 120 kDa

false

Sanger Sequencing - Human ADAM17 knockout HCT116 cell line (AB266873)
  • Sanger seq

Unknown

Sanger Sequencing - Human ADAM17 knockout HCT116 cell line (AB266873)

Homozygous : 1 bp deletion in exon5

关键信息

细胞类型

HCT116

种属

Human

组织

Colon

形式

Liquid

form

敲除验证

Sanger Sequencing

突变描述

Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp deletion in exon 5

抗生素抗性

Puromycin 1µg/mL

疾病

Carcinoma

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搭配使用产品

Cell Lines & Lysates

AB280045

Human ADAM17 knockout K-562 cell line

Western blot - Human ADAM17 knockout K-562 cell line (AB280045)

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  • 0
Cell Lines & Lysates

AB266015

Human DNAJC10 knockout HeLa cell line

Sanger Sequencing - Human DNAJC10 knockout HeLa cell line (AB266015)

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  • 0
Primary Antibodies

AB210655

Alexa Fluor® 594 Anti-Integrin alpha 2 antibody [EPR17338] - C-terminal

RabMAb|Recombinant|KO Validated

Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 594 Anti-Integrin alpha 2 antibody [EPR17338] - C-terminal (AB210655)

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  • 0
Proteins & Peptides

AB101661

Recombinant human HDAC1 protein (GST tag C-Terminus)

Functional Studies - Recombinant human HDAC1 protein (AB101661)

  • 0
  • 0
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反应性数据

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" }, "Sanger seq": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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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.

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规格

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性能和储存信息

基因名称
ADAM17
基因编辑类型
Knockout
基因编辑方法
CRISPR technology
敲除验证
Sanger Sequencing
合子性
Homozygous
运输条件
Dry Ice
推荐的短期储存条件
-196°C
推荐的长期储存条件
-196°C
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补充信息

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

ADAM17 also known as TACE (tumor necrosis factor-alpha converting enzyme) is a metalloprotease belonging to the ADAM (a disintegrin and metalloprotease) family. This protein functions by cleaving and shedding extracellular portions of cell surface proteins. It weighs approximately 93 kDa. ADAM17 is expressed in a range of tissues including the heart liver kidney and brain. The protein plays a role in the regulation of various biological processes through the activation and inactivation of membrane-bound precursor proteins.
Biological function summary

The ADAM17 protein regulates a multitude of cellular responses such as inflammation and growth factor signaling. It is not part of a complex but interacts with various substrates in the cell membrane. ADAM17 cleaves precursors to release soluble cytokines and growth factors like tumor necrosis factor-alpha (TNF-alpha) and epidermal growth factor receptor (EGFR) ligands which modulate cell proliferation migration and apoptosis.

Pathways

ADAM17 plays an integral role in both the TNF and EGFR signaling pathways. These pathways are important in maintaining normal cellular homeostasis. ADAM17 interacts with proteins like TNF receptors and ligands of the EGFR family integrating signaling that affects immune response and cell growth. The regulatory function of ADAM17 in these pathways makes it an important target for modulating cellular responses triggered by external stimuli.

ADAM17 has significant implications in conditions such as rheumatoid arthritis and cancer. In rheumatoid arthritis the protein's shedding activity influences inflammatory cytokines contributing to the disease's pathogenesis alongside proteins like TNF-alpha. In cancer ADAM17's ability to release EGFR ligands affects tumor cell proliferation and metastasis with interactions involving EGFR proteins. It is a target of therapeutic interest with research focused on developing inhibitors to manage these diseases effectively.
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质量控制

STR 分析

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

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细胞培养

生物安全等级

EU: 1 US: 1

贴壁/悬浮

Adherent

性别

Male

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. <p>1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.<br>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.<br>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.<br>4. Incubate the culture at 37°C incubator with 5% CO<sub>2</sub>. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.<br>5. Once confluent passage into an appropriate flask at a density of 2x10<sup>4</sup> cells/cm<sup>2</sup>. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.</p>

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.

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产品实验方案

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靶点信息

See full target information ADAM17
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