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

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Images

Western blot - Human DUSP4 knockout A549 cell line [F7] (AB319090), expandable thumbnail
  • Next Generation Sequencing - Human DUSP4 knockout A549 cell line [F7] (AB319090), expandable thumbnail

Key facts

Cell type

A549

Species or organism

Human

Tissue

Lung

Form

Liquid

Knockout validation

Next Generation Sequencing, Western blot

Mutation description

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

Recommended products

DUSP4 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9,109 bp deletion (allele 1) and 110 bp deletion (allele 2) in exon 2, CCDS6072.1.

Key facts

Cell type

A549

Form

Liquid

Mutation description

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

Disease

Carcinoma

Concentration
Loading...

Properties

Gene name

DUSP4

Gene editing type

Knockout

Gene editing method

CRISPR technology

Knockout validation

Next Generation Sequencing, Western blot

Quality control

STR analysis

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

Cell culture

Biosafety level

EU: 1 US: 1

Viability

~ 80%

Adherent/suspension

Adherent

Gender

Male

Handling procedures

Initial handling guidelines

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.

Subculture guidelines

  • 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.

Culture medium

F-12K + 10% FBS

Cryopreservation medium

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

Storage

Shipped at conditions

Dry Ice

Appropriate short-term storage duration

A few days

Appropriate short-term storage conditions

-196°C

Appropriate long-term storage conditions

-196°C

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

2 product images

  • Western blot - Human DUSP4 knockout A549 cell line [F7] (ab319090), expandable thumbnail

    Western blot - Human DUSP4 knockout A549 cell line [F7] (ab319090)

    Western blot: Anti-DUSP4 antibody [EPR19881] (Anti-DUSP4 antibody [EPR19881] ab216576) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (Anti-Calnexin antibody [CANX/1543] ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-DUSP4 antibody [EPR19881] 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] (Anti-DUSP4 antibody [EPR19881] ab216576) 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

    Performed under reducing conditions.

    Observed band size: 40 kDa

  • Next Generation Sequencing - Human DUSP4 knockout A549 cell line [F7] (ab319090), expandable thumbnail

    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

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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