Human RHOA knockout HEK-293T cell line
- Advanced Validation
- What is this?
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RHOA KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 4 bp insertion in exon 2 and Insertion of the selection cassette in exon 2.
View Alternative Names
ARH12, ARHA, Aplysia ras related homolog 12, Oncogene RHO H12, RHO H12, RHO12, RHOA_HUMAN, Ras homolog family member A, Ras homolog gene family member A, Rho cDNA clone 12, Small GTP binding protein Rho A, Transforming protein Rho A
- WB
Lab
Western blot - Human RHOA knockout HEK-293T cell line (AB266592)
Lanes 1- 4 : Merged signal (red and green). Green - ab187027 observed at 21 kDa. Red - Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) observed at 50 kDa.
ab187027 was shown to react with RhoA in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266592 (knockout cell lysate ab257637) was used. Wild-type HEK-293T and RHOA knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab187027 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) overnight at 4° at a 1 in 5000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes:
Western blot - Anti-RhoA antibody [EPR18134] (<a href='/en-us/products/primary-antibodies/rhoa-antibody-epr18134-ab187027'>ab187027</a>) at 1/5000 dilution
Lanes 1 and 3:
Wild-type HEK-293T cell lysate at 20 µg
Lanes 2 and 4:
RHOA knockout HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human RHOA knockout HEK-293T cell line (ab266592)
Predicted band size: 22 kDa
Observed band size: 21 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human RHOA knockout HEK-293T cell line (AB266592)
Allele-2 : Insertion of the selection cassette in exon 2.
- Sanger seq
Unknown
Sanger Sequencing - Human RHOA knockout HEK-293T cell line (AB266592)
Allele-1 : 4 bp insertion in exon 2
Reactivity data
Product details
Recommended control: Human wild-type HEK293T cell line (ab255449). 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, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and 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
- 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
The role of RhoA extends to cell proliferation and differentiation. It is a component of the Rho-GTPase cycle interacting with numerous effectors to transmit signals from the cell membrane to the actin cytoskeleton. RhoA influences the assembly of stress fibers and focal adhesions. It plays a part in cellular adhesion and migration making it essential for development and wound healing processes. As a central player in these activities RhoA affects several cellular responses to external stimuli.
Pathways
Research places RhoA prominently in the regulation of the Rho-ROCK (Rho-associated protein kinase) pathway and the planar cell polarity pathway. The Rho-ROCK pathway contributes to cytoskeletal dynamics and cell contractility while the planar cell polarity pathway influences tissue architecture during embryonic development. RhoA interacts with G17A in these pathways coordinating activities such as smooth muscle contraction and neuronal growth. These interactions also involve cross-talk with other signals linked to GTPases like Rac1 and Cdc42.
Quality control
STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
Biosafety level
EU: 2 US: 2
Adherent/suspension
Adherent
Gender
Female
Target data
Complementary Products
![Western blot - Anti-RhoA antibody [EPR18134] (AB187027)](https://content.abcam.com/products/images/rhoa-antibody-epr18134-ab187027--western-blot-img2083.jpg)
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