Human KLF5 knockout A549 cell line
- Advanced Validation
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KLF5 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout.
View Alternative Names
BTE-binding protein 2, BTEB 2, Basic transcription element-binding protein 2, Colon krueppel-like factor, GC-box-binding protein 2, IKLF, Intestinal-enriched krueppel-like factor, KLF5_HUMAN, Krueppel-like factor 5, Kruppel like factor 5 intestinal, Transcription factor BTEB 2
- WB
Lab
Western blot - Human KLF5 knockout A549 cell line (AB301031)
Western blot : Anti-KLF5 antibody [BLR243L] (ab314109) 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, ab314109 was shown to bind specifically to KLF5. A band was observed at 55 kDa in wild-type A549 cell lysates with no signal observed at this size in KLF5 knockout cell line. To generate this image, wild-type and KLF5 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-KLF5 antibody [BLR243L] - BSA free (<a href='/en-us/products/primary-antibodies/klf5-antibody-blr243l-bsa-free-ab314109'>ab314109</a>) at 1/1000 dilution
Lane 1:
Wild-type A549 cell lysate at 20 µg
Lane 2:
KLF5 knockout A549 cell lysate at 20 µg
Lane 3:
HAP1 cell lysate at 20 µg
Lane 4:
OVCAR-3 cell lysate at 20 µg
Lane 5:
PANC-1 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: 55 kDa
false
- WB
Lab
Western blot - Human KLF5 knockout A549 cell line (AB301031)
Western blot : Anti-KLF5 antibody (ab137676) staining at 1/2000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab137676 was shown to bind specifically to KLF5. A band was observed at 55 kDa in wild-type A549 cell lysates with no signal observed at this size in KLF5 knockout cell line. To generate this image, wild-type and KLF5 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-KLF5 antibody (<a href='/en-us/products/primary-antibodies/klf5-antibody-ab137676'>ab137676</a>) at 1/2000 dilution
Lane 1:
Wild-type A549 cell lysate at 20 µg
Lane 2:
KLF5 knockout A549 cell lysate at 20 µg
Lane 3:
HAP1 cell lysate at 20 µg
Lane 4:
OVCAR-3 cell lysate at 20 µg
Lane 5:
PANC-1 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: 55 kDa
false
- NGS
Lab
Next Generation Sequencing - Human KLF5 knockout A549 cell line (AB301031)
4 bp deletion after Ala 116 of WT protein, 55 bp deletion downstream of stop codon.
- WB
Lab
Western blot - Human KLF5 knockout A549 cell line (AB301031)
Western blot : Anti-KLF5 antibody ab137676 staining at 1/2000 dilution, shown in green; Mouse anti GAPDH (ab8245) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 55 kDa in Wild-type HCT 116 cell lysates with no signal observed at this size in KLF5 knockout HCT 116 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc 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 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.
All lanes:
Western blot - Anti-KLF5 antibody (<a href='/en-us/products/primary-antibodies/klf5-antibody-ab137676'>ab137676</a>) at 1/2000 dilution
Lane 1:
Wild-type HCT 116 at 20 µg
Lane 2:
Western blot - Human KLF5 knockout HCT116 cell line (<a href='/en-us/products/cell-lines/human-klf5-knockout-hct116-cell-line-ab301030'>ab301030</a>) at 20 µg
Lane 3:
Western blot - Human wild-type A549 cell line (ab288558) at 20 µg
Lane 4:
Western blot - Human KLF5 knockout A549 cell line (ab301031) at 20 µg
Secondary
Lanes 1 - 4:
Goat anti-Rabbit 800CW at 1/20000 dilution
Lanes 1 - 4:
Goat anti-Mouse 680RD at 1/20000 dilution
Predicted band size: 51 kDa,55 kDa
Observed band size: 55 kDa,37 kDa
false
- WB
Lab
Western blot - Human KLF5 knockout A549 cell line (AB301031)
Western blot : Anti-KLF5 antibody [BLR243L] - BSA free ab314109 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH (ab8245) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 55 kDa in Wild-type HCT 116 cell lysates with no signal observed at this size in KLF5 knockout HCT 116 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc 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 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.
All lanes:
Western blot - Anti-KLF5 antibody [BLR243L] - BSA free (<a href='/en-us/products/primary-antibodies/klf5-antibody-blr243l-bsa-free-ab314109'>ab314109</a>) at 1/1000 dilution
Lane 1:
Wild-type HCT 116 at 20 µg
Lane 2:
Western blot - Human KLF5 knockout HCT116 cell line (<a href='/en-us/products/cell-lines/human-klf5-knockout-hct116-cell-line-ab301030'>ab301030</a>) at 20 µg
Lane 3:
Western blot - Human wild-type A549 cell line (ab288558) at 20 µg
Lane 4:
Western blot - Human KLF5 knockout A549 cell line (ab301031) at 20 µg
Secondary
Lanes 1 - 4:
Goat anti-Rabbit 800CW at 1/20000 dilution
Lanes 1 - 4:
Goat anti-Mouse 680RD at 1/20000 dilution
Predicted band size: 51 kDa,55 kDa
Observed band size: 55 kDa,37 kDa
false
Reactivity data
Product details
Although we aim to provide customers with a homozygous clone, feasibility will be dependent on the biology of the protein. Should only heterozygous edits be achieved, you will be notified of the outcome and be asked to confirm whether the cell line is acceptable. All clones will be accompanied with DNA sequencing data, and the mutation description.
Recommended control: Human wild-type A549 cell line (ab288558). 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.
What's included?
Properties and storage information
Gene name
Gene editing type
Gene editing method
Knockout validation
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
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.
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
Kruppel-like factor 5 modulates gene transcription by binding to GC-rich elements in promoter regions influencing various cellular processes. KLF5 can interact with other transcription factors and co-regulators forming complexes important for its function. It significantly impacts epithelial cell dynamics affecting cell transformation differentiation and migration. In particular it plays a pivotal role in maintaining the proliferative capacity of epithelial cells and supporting wound healing processes.
Pathways
Kruppel-like factor 5 holds a notable position in the TGF-beta and Wnt signaling pathways which govern cellular growth control and development. In the TGF-beta pathway KLF5 can antagonize SMAD-mediated transcription suppression promoting cell proliferation particularly in epithelial stem cells. In the Wnt pathway it influences beta-catenin activity essential for epithelial cell renewal and maintenance. Through these pathways KLF5 interacts with proteins such as SMAD3 and beta-catenin coordinating cell turnover and tissue homeostasis.
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Target data
Complementary Products
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
For licensing inquiries, please contact partnerships@abcam.com