Human ERBB3 knockout MCF7 cell line
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- NGS
Lab
Next Generation Sequencing - Human ERBB3 knockout MCF7 cell line (AB286320)
101 bp deletion in exon 2, CCDS31833.1
- WB
Lab
Western blot - Human ERBB3 knockout MCF7 cell line (AB286320)
Western blot : Anti-ErbB3 / HER3 antibody [EPR22669-25] ab255607 staining at 1/1000 dilution, shown in green; Mouse anti-CANX (ab238078) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 90-175 kDa in Wild-type MCF7 UNBOILED cell lysates with no signal observed at this size in ERBB3 knockout MCF7 UNBOILED cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5pc BSA 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-ErbB3 / HER3 antibody [EPR22669-25] (<a href='/en-us/products/primary-antibodies/erbb3-her3-antibody-epr22669-25-ab255607'>ab255607</a>) at 1/1000 dilution
Lane 1:
Wild-type MCF7 UNBOILED at 20 µg
Lane 2:
Western blot - Human ERBB3 knockout MCF7 cell line (ab286320) at 20 µg
Lane 3:
U-2 OS UNBOILED at 20 µg
Lane 4:
Raji UNBOILED 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: 148 kDa,185 kDa
Observed band size: 90-175 kDa,76 kDa
false
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Proteins & Peptides
AB307190
Recombinant Human ErbB3 / HER3 protein (Fc Chimera) (Active)
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Reactivity data
Product details
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
HER3 is essential for activating important signaling pathways that regulate proliferation differentiation and survival. The ErbB3 protein forms heterodimers with other ErbB family members such as ErbB2 (HER2) which is necessary for its signaling capability. The HER3 protein is abundant in heart and nervous tissues indicating its role in cardiovascular and neurological functions. Its activation modulates gene expression through downstream effectors mediated by dimerization.
Pathways
HER3 plays a pivotal role in the PI3K/Akt and MAPK signaling pathways. These pathways are fundamental in cellular responses like growth and survival. HER3 serves as a major partner for HER2 in these signaling cascades enhancing downstream signaling effects. HER3's connection to these pathways makes it a critical node collaborating closely with proteins such as HER2 and EGFR to maintain efficient communication and function within these networks.
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