Human Arhgef1 (p115-RhoGEF) knockout HEK-293T cell line
- Advanced Validation
- What is this?
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ARHGEF1 KO cell line available to order. KO validated. Free of charge wild type control available. Knockout achieved by using CRISPR/Cas9, Homozygous: 2 bp deletion in exon 3. To order both knockout and wild-type control cells: select '2 x 1000000 Cells/vial'. To order only knockout cells: select '1000000 Cells/vial'.
View 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
- WB
Lab
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] (ab7291) loading control staining at 1/20000 dilution shown in red. In Western blot 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 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
All lanes:
Western blot - Anti-p115-RhoGEF antibody (<a href='/en-us/products/primary-antibodies/p115-rhogef-antibody-ab223759'>ab223759</a>) 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
Predicted band size: 102 kDa
Observed band size: 85 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human Arhgef1 (p115-RhoGEF) knockout HEK-293T cell line (AB266667)
Homozygous : 2 bp deletion in exon3
- Cell Culture
Unknown
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.
Reactivity data
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
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.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Complementary Products
Product promise
Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.
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