Human RHOG knockout HEK-293T cell line
- Advanced Validation
- What is this?
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RHOG KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, Homozygous: 1 bp insertion in exon 2.
View Alternative Names
ARHG, RHOG_HUMAN, Ras homolog family member G, Rho related GTP binding protein RhoG precursor, Rho-related GTP-binding protein RhoG
- WB
Lab
Western blot - Human RHOG knockout HEK-293T cell line (AB266753)
False colour image of Western blot : Anti-RHOG antibody [1F3-RHOG] staining at 1/1000 dilution shown in green; Rabbit anti-alpha Tubulin antibody [EP1332Y] (ab52866) loading control staining at 1/20000 dilution shown in red. In Western blot ab254147 was shown to bind specifically to RHOG. A band was observed at 18 kDa in wild-type HEK-293T cell lysates with no signal observed at this size in RHOG knockout cell line ab266753 (knockout cell lysate ab263328). To generate this image wild-type and RHOG 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-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) at 1/20000 dilution.
All lanes:
Western blot - Anti-RHOG antibody [1F3-RHOG] (<a href='/en-us/products/primary-antibodies/rhog-antibody-1f3-rhog-ab254147'>ab254147</a>) at 1/1000 dilution
Lane 1:
Wild-type HEK-293T cell lysate at 20 µg
Lane 2:
RHOG knockout HEK-293T cell lysate at 20 µg
Lane 2:
Western blot - Human RHOG knockout HEK-293T cell line (ab266753)
Lane 3:
Jurkat cell lysate at 20 µg
Lane 4:
SH-SY5Y cell lysate at 20 µg
Predicted band size: 21 kDa
Observed band size: 18 kDa
false
- Sanger seq
Unknown
Sanger Sequencing - Human RHOG knockout HEK-293T cell line (AB266753)
Homozygous : 1 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
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
RHOG regulates cell shape and motility. It is involved in forming membrane protrusions such as filopodia and lamellipodia. RHOG can act as a part of a protein complex with other GTPases. It controls actin cytoskeleton dynamics by activating pathways needed for cell migration and chemotaxis. This protein significantly aids immune cell function during immune surveillance and inflammatory responses.
Pathways
RHOG interacts with diverse signaling cascades. It activates the Rac1 pathway influencing cellular migration and growth processes. RHOG collaborates with proteins like PI3-kinase in these pathways contributing to the regulation of cytoskeletal organization. RHOG involvement in these signaling networks ensures cells respond aptly to external cues maintaining cellular integrity and function.
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
![Immunocytochemistry/ Immunofluorescence - Anti-RHOG antibody [1F3-RHOG] (AB254147)](https://content.abcam.com/products/images/rhog-antibody-1f3-rhog-ab254147--immunocytochemistry-immunofluorescence-img65910.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