ARHGEF1 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 2 bp deletion in exon 3.
Images
Key facts
- Cell type
- HEK-293T
- Species or organism
- Human
- Tissue
- Kidney
- Form
- Liquid
- Knockout validation
- Sanger Sequencing
- Mutation description
- Knockout achieved by using CRISPR/Cas9, Homozygous: 2 bp deletion in exon 3
Alternative names
115 kD protein, 115 kDa guanine nucleotide exchange factor, ARHG1_HUMAN, ARHGEF 1, GEF 1, LBCL 2, LSC, Lsc homolog, Rho guanine nucleotide exchange factor (GEF) 1, Rho guanine nucleotide exchange factor 1, Sub1.5, p115-RhoGEF, p115RhoGEF
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ARHGEF1 KO cell line available to order. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 2 bp deletion in exon 3.
Key facts
- Cell type
- HEK-293T
- Species or organism
- Human
- Tissue
- Kidney
- Form
- Liquid
- Knockout validation
- Sanger Sequencing
- Mutation description
- Knockout achieved by using CRISPR/Cas9, Homozygous: 2 bp deletion in exon 3
- Concentration
Properties
- Gene name
- ARHGEF1
- Gene editing type
- Knockout
- Gene editing method
- CRISPR technology
- Knockout validation
- Sanger Sequencing
- Zygosity
- Homozygous
Quality control
- STR analysis
- CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
- EU: 2 US: 2
- 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 p115-RhoGEF also known as ARHGEF1 is a guanine nucleotide exchange factor that primarily activates RhoA a small GTPase. It has a molecular mass of approximately 115 kDa. You find p115-RhoGEF predominantly expressed in the heart lung and brain tissues. This protein facilitates the exchange of GDP for GTP on RhoA triggering downstream signaling events essential for cellular functions like growth motility and organization of cytoskeletal structures.
- Biological function summary
P115-RhoGEF plays an important role in regulating actin cytoskeleton dynamics and transmitting signals from G-protein-coupled receptors. The protein is part of a signaling complex that includes heterotrimeric G proteins. Through its influence on the actin cytoskeleton p115-RhoGEF helps control changes in cell shape and movement. These activities contribute significantly to various cellular processes including differentiation and migration.
- Pathways
P115-RhoGEF is an essential component in the RhoA signaling pathway. This pathway regulates cytoskeletal dynamics and has critical implications for processes like cell adhesion and migration. p115-RhoGEF collaborates with other proteins such as Gα12/13 subunits to mediate its signaling effects. Besides the RhoA pathway p115-RhoGEF also interacts with the PI3K/Akt pathway influencing cellular survival and apoptosis.
- Associated diseases and disorders
Disruptions in p115-RhoGEF activity link to cardiovascular diseases and certain forms of cancer. Alterations in its function may lead to improper cell movement contributing to metastasis in cancer development. Additionally abnormal regulation of RhoA signaling connected to p115-RhoGEF could compromise vascular integrity implicating this protein in hypertension. During disease progression p115-RhoGEF interacts with other proteins like ROCK1 to exert these pathological effects.
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3 product images
Western blot - Human Arhgef1 (p115-RhoGEF) knockout HEK-293T cell line (ab266667)
False colour image of Western blot: Anti-p115-RhoGEF antibody staining at 0.04 µg/ml 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-p115-RhoGEF antibody ab223759 was shown to bind specifically to p115-RhoGEF. A band was observed at 85 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in Arhgef1 knockout cell line ab266667 (knockout cell lysate Human Arhgef1 (p115-RhoGEF) knockout HEK-293T cell lysate ab257842). To generate this image wild-type and Arhgef1 knockout HEK-293T 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°. 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 (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.
All lanes: Western blot - Anti-p115-RhoGEF antibody (Anti-p115-RhoGEF antibody ab223759) at 0.04 µg/mL
Lane 1: Wild-type HEK-293T Scraped cell lysate at 20 µg
Lane 2: ARHGEF1 knockout HEK-293T Scraped cell lysate at 20 µg
Lane 2: Western blot - Human Arhgef1 (p115-RhoGEF) knockout HEK-293T cell line (ab266667)
Lane 3: HeLa cell lysate at 20 µg
Lane 4: U-2 OS cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 102 kDa
Observed band size: 85 kDa
Cell Culture - Human Arhgef1 (p115-RhoGEF) knockout HEK-293T cell line (ab266667)
Representative images of Arhgef1 knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS XL Core microscope.
Sanger Sequencing - Human Arhgef1 (p115-RhoGEF) knockout HEK-293T cell line (ab266667)
Homozygous: 2 bp deletion in exon3
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