Human IGF1R knockout MCF7 cell line
- Advanced Validation
- What is this?
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(0 Publication)
- WB
Lab
Western blot - Human IGF1R knockout MCF7 cell line (AB287507)
False colour image of Western blot : Anti-IGF1 Receptor antibody [EPR19322] staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab182408 was shown to bind specifically to IGF1 Receptor. A band was observed at 82 kDa (beta chain) in wild-type MCF7 cell lysates with no signal observed at this size in IGF1R knockout cell line. To generate this image, wild-type and IGF1R knockout MCF7 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution 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-IGF1 Receptor antibody [EPR19322] (<a href='/en-us/products/primary-antibodies/igf1-receptor-antibody-epr19322-ab182408'>ab182408</a>) at 1/1000 dilution
Lane 1:
Wild-type MCF7 cell lysate at 20 µg
Lane 2:
IGF1R knockout MCF7 cell lysate at 20 µg
Lane 3:
HeLa cell lysate at 20 µg
Lane 4:
HDLM-2 cell lysate at 20 µg
Observed band size: 82 kDa
false
- WB
Lab
Western blot - Human IGF1R knockout MCF7 cell line (AB287507)
False colour image of Western blot : Anti-IGF1 Receptor antibody [EPR23027-204] staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab263903 was shown to bind specifically to IGF1 Receptor. A band was observed at 105-125 kDa (alpha chain) in wild-type MCF7 cell lysates with no signal observed at this size in IGF1R knockout cell line. To generate this image, wild-type and IGF1R knockout MCF7 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution 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-IGF1 Receptor antibody [EPR23027-204] (<a href='/en-us/products/primary-antibodies/igf1-receptor-antibody-epr23027-204-ab263903'>ab263903</a>) at 1/1000 dilution
Lane 1:
Wild-type MCF7 cell lysate at 20 µg
Lane 2:
IGF1R knockout MCF7 cell lysate at 20 µg
Lane 3:
HeLa cell lysate at 20 µg
Lane 4:
HDLM-2 cell lysate at 20 µg
Observed band size: 105-125 kDa
false
- WB
Lab
Western blot - Human IGF1R knockout MCF7 cell line (AB287507)
False colour image of Western blot : Anti-IGF1 Receptor antibody [EPR23027-80] staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab263907 was shown to bind specifically to IGF1 Receptor. A band was observed at 105-125 kDa (alpha chain) in wild-type MCF7 cell lysates with no signal observed at this size in IGF1R knockout cell line. To generate this image, wild-type and IGF1R knockout MCF7 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution 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-IGF1 Receptor antibody [EPR23027-80] (<a href='/en-us/products/primary-antibodies/igf1-receptor-antibody-epr23027-80-ab263907'>ab263907</a>) at 1/1000 dilution
Lane 1:
Wild-type MCF7 cell lysate at 20 µg
Lane 2:
IGF1R knockout MCF7 cell lysate at 20 µg
Lane 3:
HeLa cell lysate at 20 µg
Lane 4:
HDLM-2 cell lysate at 20 µg
Observed band size: 105-125 kDa
false
- NGS
Lab
Next Generation Sequencing - Human IGF1R knockout MCF7 cell line (AB287507)
182 bp deletion after the Ile55 of the WT protein
Complementary Products
Primary Antibodies
AB182408
Anti-IGF1 Receptor antibody [EPR19322]
RabMAb|Recombinant|KO Validated|20ul selling size
![Immunocytochemistry/ Immunofluorescence - Anti-IGF1 Receptor antibody [EPR19322] (AB182408)](https://content.abcam.com/products/images/igf1-receptor-antibody-epr19322-ab182408--immunocytochemistry-immunofluorescence-img58812.jpg)
- 4
- 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 MCF7 cell line (ab288560). 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
- Slow to trypsinise.
- All seeding densities should be based on cell counts gained by established methods.
- A guide seeding density of 5-7x104 cells/cm2 is recommended.
- Cells should be passaged when they have achieved 80-90% confluence.
Culture medium
MEM + 10% FBS + 0.01 mg/ml bovine insulin
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
IGF1R contributes to several cellular processes including cell proliferation differentiation and survival. This receptor forms a complex upon ligand binding and undergoes autophosphorylation to activate intracellular signaling cascades. IGF1R activation recruits and phosphorylates insulin receptor substrates enabling the downstream signaling pathways that promote cell growth and survival. Its function in cellular responses makes it a focal point in understanding cell biology.
Pathways
IGF1R holds a central position in the PI3K/AKT and MAPK signaling cascades. These pathways play significant roles in cellular growth proliferation and survival. IGF1R phosphorylates various downstream effectors such as IRS-1 and Shc linking it to the activation of AKT and ERK respectively. Therefore it shares pathways with related proteins like the insulin receptor highlighting its importance in mediating similar biological responses.
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Female
Target data
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