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AB319090

人DUSP4 knockout A549 cell line [F7]

Human DUSP4 knockout A549 cell line [F7]

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DUSP4 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9,109 bp deletion (allele

  1. and 110 bp deletion (allele 2) in exon 2, CCDS6072.1. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
2 Images
Western blot - Human DUSP4 knockout A549 cell line [F7] (AB319090)
  • WB

Lab

Western blot - Human DUSP4 knockout A549 cell line [F7] (AB319090)

Western blot : Anti-DUSP4 antibody [EPR19881] (ab216576) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab216576 was shown to bind specifically to DUSP4. A band was observed at 40 kDa in wild-type A549 cell lysates with no signal observed at this size in DUSP4 knockout cell line. To generate this image, wild-type and DUSP4 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-DUSP4 antibody [EPR19881] (<a href='/products/primary-antibodies/dusp4-antibody-epr19881-ab216576'>ab216576</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lanes 2 - 3:

DUSP4 knockout A549 cell lysate at 20 µg

Lane 4:

DUSP4 knockout A549 H15 cell lysate at 20 µg

Lane 5:

HCT 116 cell lysate at 20 µg

Lane 6:

MOLT-4 cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 40 kDa

false

Next Generation Sequencing - Human DUSP4 knockout A549 cell line [F7] (AB319090)
  • NGS

Lab

Next Generation Sequencing - Human DUSP4 knockout A549 cell line [F7] (AB319090)

109 bp deletion (allele 1) and 110 bp deletion (allele 2) in exon 2, CCDS6072.1

关键信息

细胞类型

A549

种属

Human

组织

Lung

形式

Liquid

form

敲除验证

Next Generation Sequencing,Western blot

突变描述

Knockout achieved by using CRISPR/Cas9,109 bp deletion (allele 1) and 110 bp deletion (allele 2) in exon 2, CCDS6072.1

疾病

Carcinoma

反应性数据

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

规格

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性能和储存信息

基因名称
DUSP4
基因编辑类型
Knockout
基因编辑方法
CRISPR technology
敲除验证
Next Generation Sequencing, Western blot
运输条件
Dry Ice
推荐的短期储存时间
A few days
推荐的短期储存条件
-196°C
推荐的长期储存条件
-196°C

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.

质量控制

STR 分析

D3S1358, TH01, D21S11, D18S51, D5S818, D13S317, D7S820, D16S539, CSF1PO, D8S1179, TPOX, FGA, D19S433, D2S1338, VWA

细胞培养

生物安全等级

EU: 1 US: 1

贴壁/悬浮

Adherent

性别

Male

产品实验方案

Abcam Product Promise

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