PAX6 KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control provided.
Images
Key facts
- Cell type
- A549
- Species or organism
- Human
- Tissue
- Lung
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing
Alternative names
AN, AN 2, Aniridia type II protein, D11S812E, FVH1, KIAA0552, LZTS3, Leucine zipper putative tumor suppressor 3, MGC17209, MGDA, Oculorhombin, PAX6_HUMAN, PROSAPIP1, Paired Box Gene 6, Paired box 6, Paired box gene 6 (aniridia keratitis), Paired box homeotic gene 6, Paired box protein Pax-6, ProSAP-interacting protein 1, Sey, WAGR
Recommended products
PAX6 KO cell line available to order. KO validated by Next Generation Sequencing. Free of charge wild type control provided.
Key facts
- Cell type
- A549
- Species or organism
- Human
- Tissue
- Lung
- Form
- Liquid
- Knockout validation
- Next Generation Sequencing
- Disease
- Carcinoma
- Concentration
Properties
- Gene name
- PAX6
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Next Generation Sequencing
Quality control
- STR analysis
- D13S317, D7S820, D5S818, TH01, D16S539, TPOX, CSF1PO
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
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.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
The PAX6 protein also known as paired box 6 is a transcription factor involved in the development of eyes and neural tissues. It consists of a paired domain and a homeodomain collectively giving a molecular weight of approximately 48 kDa. This protein is highly expressed in developing ocular and central nervous tissues where it controls the expression of genes necessary for proper development. PAX6 is a widely studied marker in developmental biology due to its critical role in cellular differentiation and patterning processes.
- Biological function summary
PAX6 acts as an important regulator in eye and neural development functioning alone and as part of larger complexes with other transcription factors. This role ensures the correct formation of ocular structures including the lens retina and cornea and neural elements by activating downstream targets important for cell fate decisions. The PAX6 protein intricately influences the genetic circuits that guide stem cell differentiation highlighting its importance in embryogenesis and organogenesis.
- Pathways
PAX6 is pivotal in the Wnt signaling pathway and retinoic acid signaling. Its interaction within the Wnt pathway orchestrates vital developmental processes including those of eye morphogenesis and brain patterning. The protein collaborates with other transcriptional regulators such as SOX2 and EYA1 within these pathways facilitating the complex network of signals necessary for organ development and cellular identity specification.
- Associated diseases and disorders
PAX6 mutations have strong associations with aniridia and congenital eye malformations. Mutations in PAX6 disrupt the transcriptional regulation required for eye development often resulting in severe vision defects and structural anomalies. The intracacy of its involvement extends to disorders like Wilms' tumor wherein its altered expression or function in conjunction with proteins such as WT1 may contribute to oncogenesis. Scientists frequently use PAX6 staining to investigate these conditions and better understand their molecular underpinnings.
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.
4 product images
Western blot - Human PAX6 knockout A549 cell line (ab288955)
Western blot: Anti-PAX6 antibody [EPR3352(2)] (Anti-PAX6 antibody [EPR3352(2)] ab109233) staining at 1/5000 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-PAX6 antibody [EPR3352(2)] ab109233 was shown to bind specifically to PAX6. A band was observed at 50 kDa in wild-type A549 cell lysates with no signal observed at this size in PAX6 knockout cell line. To generate this image, wild-type and PAX6 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 5 % 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-PAX6 antibody [EPR3352(2)] (Anti-PAX6 antibody [EPR3352(2)] ab109233) at 1/5000 dilution
Lane 1: Wild-type A549 cell lysate at 20 µg
Lane 2: PAX6 knockout A549 cell lysate at 20 µg
Lane 3: HEK-293 cell lysate at 20 µg
Lane 4: MCF7 cell lysate at 20 µg
SecondaryAll 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: 50 kDa
Western blot - Human PAX6 knockout A549 cell line (ab288955)
Western blot: Anti-PAX6 antibody [EPR15858] (Anti-PAX6 antibody [EPR15858] ab195045) 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-PAX6 antibody [EPR15858] ab195045 was shown to bind specifically to PAX6. A band was observed at 50 kDa in wild-type A549 cell lysates with no signal observed at this size in PAX6 knockout cell line. To generate this image, wild-type and PAX6 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 5 % 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-PAX6 antibody [EPR15858] (Anti-PAX6 antibody [EPR15858] ab195045) at 1/1000 dilution
Lane 1: Wild-type A549 cell lysate at 20 µg
Lane 2: PAX6 knockout A549 cell lysate at 20 µg
Lane 3: HEK-293 cell lysate at 20 µg
Lane 4: MCF7 cell lysate at 20 µg
SecondaryAll 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: 50 kDa
Western blot - Human PAX6 knockout A549 cell line (ab288955)
Western blot: Anti-PAX6 antibody (Anti-PAX6 antibody ab238527) 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-PAX6 antibody ab238527 was shown to bind specifically to PAX6. A band was observed at 50 kDa in wild-type A549 cell lysates with no signal observed at this size in PAX6 knockout cell line. To generate this image, wild-type and PAX6 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 5 % 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-PAX6 antibody (Anti-PAX6 antibody ab238527) at 1/1000 dilution
Lane 1: Wild-type A549 cell lysate at 20 µg
Lane 2: PAX6 knockout A549 cell lysate at 20 µg
Lane 3: HEK-293 cell lysate at 20 µg
Lane 4: MCF7 cell lysate at 20 µg
SecondaryAll 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: 50 kDa
Next Generation Sequencing - Human PAX6 knockout A549 cell line (ab288955)
55 bp deletion after Try157
Downloads
Product protocols
- Visit the general protocols
- Visit troubleshooting
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