Human GRB2 heterozygous knockout A-431 cell line
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GRB2 KO cell line available to order. KO validated by. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Heterozygous: 80 bp deletion in exon 2 and intron 2-3, and wild-type. To order both knockout and wild-type control cells: select 2 x 1000000Cells/vial. To order only knockout cells: select 1000000Cells/vial.
View Alternative Names
ASH, Abundant SRC homology, Adapter protein GRB2, Ash protein, EGFRBP GRB2, Epidermal growth factor receptor binding protein, Epidermal growth factor receptor binding protein GRB2, GRB2 adapter protein, GRB2_HUMAN, Grb3 3, Growth factor receptor bound protein 3, Growth factor receptor-bound protein 2, HT027, MST084, MSTP084, NCKAP2, OTTHUMP00000166096, OTTHUMP00000166097, OTTHUMP00000166098, Protein ash, SEM5, SH2/SH3 adapter GRB2
- WB
Lab
Western blot - Human GRB2 heterozygous knockout A-431 cell line (AB276085)
Anti-GRB2 antibody [Y301] (ab32111) staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab32111 was shown to bind specifically to GRB2. A band was observed at 26 kDa in wild-type A431 cell lysates with a reduction in signal observed at this size in GRB2 heterozygous knockout cell line. To generate this image, wild-type and GRB2 heterozygous knockout A431 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-GRB2 antibody [Y301] (<a href='/en-us/products/primary-antibodies/grb2-antibody-y301-ab32111'>ab32111</a>) at 1/1000 dilution
Lane 1:
Wild-type A431 cell lysate at 20 µg
Lane 2:
Western blot - Human GRB2 heterozygous knockout A-431 cell line (ab276085)
Lane 2:
GRB2 knockout A431 cell lysate at 20 µg
Lane 3:
HCT 116 cell lysate at 20 µg
Lane 4:
HeLa cell lysate at 20 µg
Secondary
Lanes 1 - 4:
Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution
Lanes 1 - 4:
Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Predicted band size: 25 kDa
Observed band size: 26 kDa
false
- WB
Lab
Western blot - Human GRB2 heterozygous knockout A-431 cell line (AB276085)
All lanes:
Western blot - Anti-GRB2 antibody [Y237] (<a href='/en-us/products/primary-antibodies/grb2-antibody-y237-ab32037'>ab32037</a>) at 1/5000 dilution
Lane 1:
Wild-type A431 cell lysate at 20 µg
Lane 2:
Western blot - Human GRB2 heterozygous knockout A-431 cell line (ab276085)
Lane 2:
GRB2 knockout A431 cell lysate at 20 µg
Lane 3:
HCT 116 cell lysate at 20 µg
Lane 4:
HeLa cell lysate at 20 µg
Secondary
Lanes 1 - 4:
Goat anti-Rabbit IgG H&L 800CW at 1/20000 dilution
Lanes 1 - 4:
Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Predicted band size: 25 kDa
Observed band size: 26 kDa
false
- Sanger seq
Supplier Data
Sanger Sequencing - Human GRB2 heterozygous knockout A-431 cell line (AB276085)
Sanger Sequencing - Human GRB2 heterozygous knockout A-431 cell line (ab276085)
80 bp deletion in exon 2 and intron 2-3; and wild-type.
Product details
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
Zygosity
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.
Culture medium
DMEM (High Glucose) + 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
GRB2 functions as an adaptor protein that connects activated receptor tyrosine kinases to intracellular pathways. It often forms complexes with other proteins such as the SOS protein which further propagates signaling cascades critical for cell proliferation and differentiation. GRB2's ability to mediate these interactions contributes to cellular responses to external stimuli including growth factors and hormones.
Pathways
GRB2 plays an important role in the Ras-MAPK signaling pathway. It interacts with proteins like SOS and Ras enabling signal transduction from the cell surface to the nucleus which is vital for processes such as cell growth and survival. GRB2 is also implicated in the PI3K-Akt pathway connecting it to another set of signaling proteins that regulate metabolism growth and survival.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 1 US: 1
Adherent/suspension
Adherent
Gender
Female
Target data
Complementary Products
![Immunocytochemistry/ Immunofluorescence - Anti-GRB2 antibody [Y237] (AB32037)](https://content.abcam.com/products/images/grb2-antibody-y237-ab32037--immunocytochemistry-immunofluorescence-img120871.jpg)
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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