GRB2 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Heterozygous: 80 bp deletion in exon 2 and intron 2-3, and wild-type.
Images
Key facts
- Cell type
- A-431
- Species or organism
- Human
- Tissue
- Skin
- Form
- Liquid
- Knockout validation
- Sanger Sequencing
- Mutation description
- Knockout achieved by using CRISPR/Cas9, Heterozygous: 80 bp deletion in exon 2 and intron 2-3, and wild-type.
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
Recommended products
GRB2 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Heterozygous: 80 bp deletion in exon 2 and intron 2-3, and wild-type.
Key facts
- Cell type
- A-431
- Species or organism
- Human
- Tissue
- Skin
- Form
- Liquid
- Knockout validation
- Sanger Sequencing
- Mutation description
- Knockout achieved by using CRISPR/Cas9, Heterozygous: 80 bp deletion in exon 2 and intron 2-3, and wild-type.
- Disease
- Epidermoid Carcinoma
- Concentration
Properties
- Gene name
- GRB2
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing
- Zygosity
- Heterozygous
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 1 US: 1
- Adherent/suspension
- Adherent
- Gender
- Female
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.
Storage
- Shipped at conditions
- Dry Ice
- Appropriate short-term storage conditions
- -196°C
- Appropriate long-term storage conditions
- -196°C
Notes
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 GRB2 protein also known as growth factor receptor-bound protein 2 is instrumental in signal transduction. GRB2 has a molecular weight of about 25 kDa. It comprises one SH2 domain and two SH3 domains which facilitate its role in linking receptor tyrosine kinases to downstream signaling molecules. GRB2 is ubiquitously expressed in various tissues signifying its importance in diverse cellular functions.
- 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.
- Associated diseases and disorders
GRB2 has significant associations with cancer and immune disorders. Aberrant activation of pathways involving GRB2 can lead to uncontrolled cell proliferation contributing to oncogenesis in different cancers. Additionally GRB2’s interactions with proteins like BCR-ABL in chronic myeloid leukemia highlight its potential role as a therapeutic target. GRB2 inhibitors could therefore offer promising avenues for treating such conditions by disrupting its pathological signaling interactions.
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3 product images
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.
Western blot - Human GRB2 heterozygous knockout A-431 cell line (ab276085)
Anti-GRB2 antibody [Y301] (Anti-GRB2 antibody [Y301] ab32111) staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-GRB2 antibody [Y301] 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] (Anti-GRB2 antibody [Y301] ab32111) 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
SecondaryLanes 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
Performed under reducing conditions.
Predicted band size: 25 kDa
Observed band size: 26 kDa
Western blot - Human GRB2 heterozygous knockout A-431 cell line (ab276085)
All lanes: Western blot - Anti-GRB2 antibody [Y237] (Anti-GRB2 antibody [Y237] ab32037) 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
SecondaryLanes 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
Performed under reducing conditions.
Predicted band size: 25 kDa
Observed band size: 26 kDa
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