Human KRT7 knockout A549 cell line (ab261867)

KRT7 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9 X = 2 bp deletion Frameshift = 100%.

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Images

Western blot - Human KRT7 knockout A549 cell line (AB261867), 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 CRISPR/Cas9 X = 2 bp deletion Frameshift = 100%

Alternative names

CK-7, Cytokeratin-7, D15Wsu77e, K2C7_HUMAN, K7, KRT7, Keratin 7, type II, Keratin type II cytoskeletal 7, Keratin, 55K type II cytoskeletal, Keratin, simple epithelial, Keratin, simple epithelial type I, K7, Keratin-7, Krt2-7, MGC11625, MGC129731, MGC3625, SCL, Sarcolectin, Type II mesothelial keratin K7, Type-II keratin Kb7

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 CRISPR/Cas9 X = 2 bp deletion Frameshift = 100%
Disease: Carcinoma
Concentration

Properties

Gene name: KRT7
Gene editing type: Knockout
Gene editing method: CRISPR technology
Knockout validation: Next Generation Sequencing, Western blot

Quality control

STR analysis: CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

Cell culture

Biosafety level: EU: 1 US: 1
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 conditions: -196°C
Appropriate long-term storage conditions: -196°C

Notes

Recommended control: Human wild-type A549 cell line (ab255450). Please note a wild-type cell line is not automatically included with a knockout cell line order, if required please add recommended wild-type cell line at no additional cost using the code WILDTYPE-TMTK1.

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.

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.

9 product images

Western blot - Human KRT7 knockout A549 cell line (ab261867)
All lanes: Western blot - Anti-Cytokeratin 7 antibody [EPR1619Y] - Cytoskeleton Marker (Anti-Cytokeratin 7 antibody [EPR1619Y] - Cytoskeleton Marker ab68459) at 1/5000 dilution
Lane 1: Wild-type A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg
Lane 2: Western blot - Human KRT7 knockout A549 cell line (ab261867) at 20 µg
Lane 3: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 4: MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg
Predicted band size: 51 kDa
Observed band size: 50 kDa
Western blot - Human KRT7 knockout A549 cell line (ab261867)
All lanes: Western blot - HRP Anti-Cytokeratin 7 antibody [EPR17078] (HRP Anti-Cytokeratin 7 antibody [EPR17078] ab209945) at 1/5000 dilution
Lane 1: Wild-type A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg
Lane 2: Western blot - Human KRT7 knockout A549 cell line (ab261867) at 20 µg
Lane 3: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 4: MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 51 kDa
Next Generation Sequencing - Human KRT7 knockout A549 cell line (ab261867)
X = 2 bp deletion
Next Generation Sequencing - Human KRT7 knockout A549 cell line (ab261867)
2 bp deletion after Arg166 of the WT protein
Western blot - Human KRT7 knockout A549 cell line (ab261867)
Western blot: Anti-KRT7 antibody [EP1620Y] (Anti-Cytokeratin 7 antibody [EP1620Y] ab68460) staining at 1/10000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-Cytokeratin 7 antibody [EP1620Y] ab68460 was shown to bind specifically to KRT7. A band was observed at 40-55 kDa in wild-type A549 cell lysates with no signal observed at this size in KRT7 knockout cell line. To generate this image, wild-type and KRT7 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-Cytokeratin 7 antibody [EP1620Y] (Anti-Cytokeratin 7 antibody [EP1620Y] ab68460) at 1/10000 dilution
Lane 1: Wild-type A549 cell lysate at 20 µg
Lane 2: KRT7 knockout A549 cell lysate at 20 µg
Lane 3: HeLa cell lysate at 20 µg
Lane 4: MCF7 cell lysate at 20 µg
Observed band size: 40-55 kDa
Western blot - Human KRT7 knockout A549 cell line (ab261867)
Western blot: Anti-KRT7 antibody [EPR17078] (Anti-Cytokeratin 7 antibody [EPR17078] - Cytoskeleton Marker ab181598) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-Cytokeratin 7 antibody [EPR17078] - Cytoskeleton Marker ab181598 was shown to bind specifically to KRT7. A band was observed at 40-55 kDa in wild-type A549 cell lysates with no signal observed at this size in KRT7 knockout cell line. To generate this image, wild-type and KRT7 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-Cytokeratin 7 antibody [EPR17078] - Cytoskeleton Marker (Anti-Cytokeratin 7 antibody [EPR17078] - Cytoskeleton Marker ab181598) at 1/10000 dilution
Lane 1: Wild-type A549 cell lysate at 20 µg
Lane 2: KRT7 knockout A549 cell lysate at 20 µg
Lane 3: HeLa cell lysate at 20 µg
Lane 4: MCF7 cell lysate at 20 µg
Observed band size: 40-55 kDa
Western blot - Human KRT7 knockout A549 cell line (ab261867)
All lanes: Western blot - HRP Anti-Cytokeratin 7 antibody [EPR1619Y] - Cytoskeleton Marker (HRP Anti-Cytokeratin 7 antibody [EPR1619Y] - Cytoskeleton Marker ab192079) at 1/5000 dilution
Lane 1: Wild-type A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg
Lane 2: Western blot - Human KRT7 knockout A549 cell line (ab261867) at 20 µg
Lane 3: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 4: MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 51 kDa
Observed band size: 51 kDa
Western blot - Human KRT7 knockout A549 cell line (ab261867)
All lanes: Western blot - Anti-Cytokeratin 7 antibody [SP52] (Anti-Cytokeratin 7 antibody [SP52] ab183344) at 1/25 dilution
Lane 1: Wild-type A549 (Human lung carcinoma cell line) whole cell lysate
Lane 2: Western blot - Human KRT7 knockout A549 cell line (ab261867)
Lane 3: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 4: MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
Performed under reducing conditions.
Predicted band size: 51 kDa
Observed band size: 50 kDa
Western blot - Human KRT7 knockout A549 cell line (ab261867)
Lanes 1 - 4: Merged signal (red and green). Green - ab119697 observed at 50 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.

ab119697 was shown to specifically react with KRT7 (Cytokeratin 7) in wild-type A549 cells as signal was lost in KRT7 knockout cell line ab261867 (knockout cell lysate Human KRT7 knockout A549 cell lysate ab261676). Wild-type and KRT7 knockout samples were subjected to SDS-PAGE. ab119697 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/25 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
All lanes: ab119697 Anti-Cytokeratin 7 antibody [SP52] at 1/25 dilution
Lane 1: Wild-type A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg
Lane 2: Western blot - Human KRT7 knockout A549 cell line (ab261867) at 20 µg
Lane 3: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 20 µg
Lane 4: MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 51 kDa
Observed band size: 50 kDa