Human GLI1 knockout A549 cell line
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GLI1 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control available. knockout. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
View Alternative Names
GLI, GLI Kruppel family member 1, GLI family zinc finger 1, GLI1_HUMAN, Glioma associated oncogene 1, Glioma associated oncogene homolog, Glioma associated oncogene homolog 1 (zinc finger protein), Glioma-associated oncogene, Oncogene GLI, Zfp 5, Zinc finger protein GLI 1
- WB
Lab
Western blot - Human GLI1 knockout A549 cell line (AB277815)
Western blot : Rabbit Monoclonal [EPR4523] to Gli1 ab134906 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 145 kDa in Wild-type A549 cell lysates with no signal observed at this size in GLI1 knockout A549 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-Gli1 antibody [EPR4523] (<a href='/en-us/products/primary-antibodies/gli1-antibody-epr4523-ab134906'>ab134906</a>) at 1/1000 dilution
Lane 1:
Wild-type A549 cell lysate at 40 µg
Lane 2:
Human GLI1 knockout A549 cell line at 20 µg
Lane 2:
Western blot - Human GLI1 knockout A549 cell line (ab277815) at 20 µg
Lane 3:
SH-SY5Y cell lysate at 40 µg
Secondary
All lanes:
Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution
Predicted band size: 118 kDa
Observed band size: 145 kDa
false
- NGS
Supplier Data
Next Generation Sequencing - Human GLI1 knockout A549 cell line (AB277815)
142 bp deletion after Leu588 (edit 1); 63 bp deletion and 1 bp insertion after Leu588 (edit 2)
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.
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
Gli1 functions to regulate cellular proliferation differentiation and survival. As part of the Gli protein family Gli1 operates in concert with other members such as Gli2 and Gli3 though it lacks significant repressor activity unlike some of its counterparts. It acts prominently in developmental processes controlling the expression of genes critical for embryogenesis. Gli1 also maintains stem cell populations and sustains tissue homeostasis in adults particularly in contexts where rapid cell division is necessary like wound healing.
Pathways
Gli1 plays a substantial role in the Hedgehog signaling pathway and interacts dynamically with this system. It receives upstream activation from the Smoothened protein which conveys signals in response to Hedgehog ligand availability. Additionally the Gli1 pathway overlaps with WNT signaling during various developmental stages. Through these interactions Gli1 coordinates tightly with other proteins to effect changes in target gene transcription that influence cell fate decisions.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Male
Target data
Complementary Products
Primary Antibodies
AB134906
RabMAb|Recombinant|KO Validated|20ul selling size
![Western blot - Anti-Gli1 antibody [EPR4523] (AB134906)](https://content.abcam.com/products/images/gli1-antibody-epr4523-ab134906--western-blot-img455335.jpg)
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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