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AB261867

Human KRT7 knockout A549 cell line

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KRT7 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control available. Knockout achieved by CRISPR/Cas9 X = 2 bp deletion Frameshift = 100%. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
9 Images
Western blot - Human KRT7 knockout A549 cell line (AB261867)
  • WB

Lab

Western blot - Human KRT7 knockout A549 cell line (AB261867)

Western blot : Anti-KRT7 antibody [EPR17078] (ab181598) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, 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 (<a href='/en-us/products/primary-antibodies/cytokeratin-7-antibody-epr17078-cytoskeleton-marker-ab181598'>ab181598</a>) 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

false

Western blot - Human KRT7 knockout A549 cell line (AB261867)
  • WB

Lab

Western blot - Human KRT7 knockout A549 cell line (AB261867)

All lanes:

Western blot - HRP Anti-Cytokeratin 7 antibody [EPR17078] (<a href='/en-us/products/primary-antibodies/hrp-cytokeratin-7-antibody-epr17078-ab209945'>ab209945</a>) 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

false

Western blot - Human KRT7 knockout A549 cell line (AB261867)
  • WB

Lab

Western blot - Human KRT7 knockout A549 cell line (AB261867)

All lanes:

Western blot - Anti-Cytokeratin 7 antibody [EPR1619Y] - Cytoskeleton Marker (<a href='/en-us/products/primary-antibodies/cytokeratin-7-antibody-epr1619y-cytoskeleton-marker-ab68459'>ab68459</a>) 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

false

Western blot - Human KRT7 knockout A549 cell line (AB261867)
  • WB

Lab

Western blot - Human KRT7 knockout A549 cell line (AB261867)

Western blot : Anti-KRT7 antibody [EP1620Y] (ab68460) staining at 1/10000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, 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] (<a href='/en-us/products/primary-antibodies/cytokeratin-7-antibody-ep1620y-ab68460'>ab68460</a>) 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

false

Next Generation Sequencing - Human KRT7 knockout A549 cell line (AB261867)
  • NGS

Lab

Next Generation Sequencing - Human KRT7 knockout A549 cell line (AB261867)

X = 2 bp deletion

Next Generation Sequencing - Human KRT7 knockout A549 cell line (AB261867)
  • NGS

Supplier Data

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)
  • WB

Lab

Western blot - Human KRT7 knockout A549 cell line (AB261867)

All lanes:

Western blot - Anti-Cytokeratin 7 antibody [SP52] (<a href='/en-us/products/primary-antibodies/cytokeratin-7-antibody-sp52-ab183344'>ab183344</a>) 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

Predicted band size: 51 kDa

Observed band size: 50 kDa

false

Western blot - Human KRT7 knockout A549 cell line (AB261867)
  • WB

Lab

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, 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 ab261676). Wild-type and KRT7 knockout samples were subjected to SDS-PAGE. ab119697 and 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 ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

<a href='/en-us/products/unavailable/cytokeratin-7-antibody-sp52-ab119697'>ab119697</a> 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

Predicted band size: 51 kDa

Observed band size: 50 kDa

false

Western blot - Human KRT7 knockout A549 cell line (AB261867)
  • WB

Lab

Western blot - Human KRT7 knockout A549 cell line (AB261867)

All lanes:

Western blot - HRP Anti-Cytokeratin 7 antibody [EPR1619Y] - Cytoskeleton Marker (<a href='/en-us/products/primary-antibodies/hrp-cytokeratin-7-antibody-epr1619y-cytoskeleton-marker-ab192079'>ab192079</a>) 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: 51 kDa

false

Key facts

Cell type

A549

Species or organism

Human

Tissue

Lung

Form

Liquid

form

Knockout validation

Next Generation Sequencing,Western blot

Mutation description

Knockout achieved by CRISPR/Cas9 X = 2 bp deletion Frameshift = 100%

Disease

Carcinoma

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Complementary Products

Cell Lines & Lysates

AB255400

Human KRT8 (Cytokeratin 8) knockout HeLa cell line

Advanced Validation

Western blot - Human KRT8 (Cytokeratin 8) knockout HeLa cell line (AB255400)

  • 0
  • 0
Primary Antibodies

AB181598

Anti-Cytokeratin 7 antibody [EPR17078] - Cytoskeleton Marker

KO Validated|RabMAb|Recombinant|Lab Essentials

Immunohistochemistry (Frozen sections) - Anti-Cytokeratin 7 antibody [EPR17078] - Cytoskeleton Marker (AB181598)

  • 5
  • 12
Primary Antibodies

AB32127

Anti-Synaptophysin antibody [YE269] - Synaptic Marker

BOND RX™ Validated|RabMAb|Recombinant|KO Validated

Western blot - Anti-Synaptophysin antibody [YE269] - Synaptic Marker (AB32127)

  • 5
  • 37
Primary Antibodies

AB15580

Anti-Ki67 antibody

BOND RX™ Validated|KO Validated

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Ki67 antibody (AB15580)

  • 5
  • 230
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Reactivity data

{ "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>" } } }
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Product details

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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What's included?

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Properties and storage information

Gene name
KRT7
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Next Generation Sequencing, Western blot
Shipped at conditions
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C
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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.

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Quality control

STR analysis

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

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Cell culture

Biosafety level

EU: 1 US: 1

Adherent/suspension

Adherent

Gender

Male

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Product protocols

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Target data

See full target information KRT7
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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

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

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