RHOA KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 4 bp insertion in exon 2 and Insertion of the selection cassette in exon 2.
Images
Key facts
- Cell type
HEK-293T
- Species or organism
Human
- Tissue
Kidney
- Knockout validation
Sanger Sequencing, Western blot
- Mutation description
Knockout achieved by using CRISPR/Cas9, 4 bp insertion in exon 2 and Insertion of the selection cassette in exon 2.
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
What's included?
Recommended products
RHOA KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 4 bp insertion in exon 2 and Insertion of the selection cassette in exon 2.
Key facts
- Cell type
HEK-293T
- Species or organism
Human
- Tissue
Kidney
- Knockout validation
Sanger Sequencing, Western blot
- Mutation description
Knockout achieved by using CRISPR/Cas9, 4 bp insertion in exon 2 and Insertion of the selection cassette in exon 2.
- Concentration
Properties
- Gene name
RHOA
- Gene editing type
Knockout
- Gene editing method
CRISPR technology
- Knockout validation
Sanger Sequencing, Western blot
Quality control
- STR analysis
CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX
Cell culture
- Biosafety level
EU: 2 US: 2
- Adherent/suspension
Adherent
- Gender
Female
Storage
- Shipped at conditions
Ambient - Can Ship with Ice
- Appropriate short-term storage conditions
-20°C
- Appropriate long-term storage conditions
-20°C
Notes
Knockout cell lysate achieved by CRISPR/Cas9.
Abcam has not and does not intend to apply for the REACH Authorisation of customers' uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
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.
Supplementary info
This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
RhoA also known as Ras homolog family member A is a small signaling GTPase. It weighs approximately 21 kDa. This protein is a part of the Rho family which includes other members like RhoB and RhoC. RhoA is widely expressed in various tissues including the brain kidney liver and lungs. Mechanically RhoA functions as a molecular switch cycling between an active GTP-bound state and an inactive GDP-bound state. This activity regulates the organization of the actin cytoskeleton and influences cell shape attachment and motility.
- 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.
- Associated diseases and disorders
RhoA has connections to cancer progression and hypertension. In cancer RhoA regulates actin dynamics influencing tumor cell migration and invasion. It forms interactions with proteins such as p53 affecting apoptotic pathways and tumor suppression. In hypertension RhoA's role in smooth muscle contraction affects blood vessel tension and blood pressure regulation. Abnormal activity in the Rho-ROCK pathway contributes to these disorders highlighting the significance of RhoA in disease mechanisms.
Product promise
We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.
In the unlikely event of one of our products not working as expected, you are covered by our product promise.
Full details and terms and conditions can be found here:
Terms & Conditions.
3 product images
Western blot - Human RHOA knockout HEK-293T cell lysate (ab257637)
Lane 1: Wild-type HEK-293T cell lysate (20µg)
Lane 2: RHOA knockout HEK-293T cell lysate (20µg)
Lane 3: Wild-type HEK-293T cell lysate (20µg)
Lane 4: RHOA knockout HEK-293T cell lysate (20µg)
Lanes 1- 4: Merged signal (red and green). Green - Anti-RhoA antibody [EPR18134] ab187027 observed at 21 kDa. Red - loading control Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 observed at 50 kDa.
Anti-RhoA antibody [EPR18134] ab187027 Anti-RhoA antibody [EPR18134] was shown to specifically react with RhoA in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line Human RHOA knockout HEK-293T cell line ab266592 (knockout cell lysate ab257637) was used. Wild-type and RhoA knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. Anti-RhoA antibody [EPR18134] ab187027 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) were incubated overnight at 4°C at 1 in 5000 and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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] (Anti-RhoA antibody [EPR18134] ab187027) 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
Performed under reducing conditions.
Predicted band size: 22 kDa
Observed band size: 21 kDa
Sanger Sequencing - Human RHOA knockout HEK-293T cell lysate (ab257637)
Allele-1: 4 bp insertion in exon 2
Sanger Sequencing - Human RHOA knockout HEK-293T cell lysate (ab257637)
Allele-2: Insertion of the selection cassette in exon 2
Downloads
Product protocols
- Visit the general protocols
- Visit troubleshooting
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