ARHGDIA KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 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, 1 bp insertion in exon 2 and Insertion of the selection cassette in exon 2.
Alternative names
ARHGDIA, GDIA 1, GDIR1_HUMAN, MGC117248, NPHS8, Rho GDI, Rho GDI 1, Rho GDP dissociation inhibitor (GDI) alpha, Rho GDP dissociation inhibitor alpha, Rho GDP-dissociation inhibitor 1, Rho-GDI alpha, RhoGDI 1, RhoGDI1, fa96g11, wu:fa96g11, zgc:55554, zgc:77681
What's included?
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ARHGDIA KO cell lysate available now. KO validated by Western blot. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, 1 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, 1 bp insertion in exon 2 and Insertion of the selection cassette in exon 2.
- Concentration
Properties
- Gene name
- ARHGDIA
- 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
RhoGDI also known as Rho GDP-dissociation inhibitor 1 is a regulatory protein with a mass of approximately 23 kDa. It primarily inhibits the dissociation of GDP from Rho GTPases such as RhoA Rac1 and Cdc42. This protein plays an important role in controlling the cycling between active GTP-bound and inactive GDP-bound states of small GTPases. RhoGDI shows expression in numerous tissues highlighting its widespread regulatory functions in cell signaling.
- Biological function summary
RhoGDI modulates the activity of Rho family GTPases—a group responsible for organizing the actin cytoskeleton which determines cell shape polarity and movement. It does not operate alone; rather it is part of a complex with membrane-bound and cytosolic proteins offering a shuttle service to balance GTPases between cellular locations. By doing so RhoGDI influences processes like cell migration and the cell cycle integral to maintaining proper cellular architecture.
- Pathways
Scientists recognize RhoGDI’s participation in key signaling pathways particularly the Rho GTPase cycle and actin cytoskeleton organization. It interacts with proteins like ROCK and PAK within these pathways to achieve precise control of cytoskeletal dynamics. RhoGDI coordinates with these proteins to play a central role in cellular responses to external stimuli impacting processes like wound healing and cellular development.
- Associated diseases and disorders
RhoGDI has associations with cancer and neurodegenerative diseases. Its deregulation can disrupt the Rho GTPase pathways leading to uncontrolled cell proliferation in cancers such as breast and prostate cancer. Additionally altered RhoGDI function affects neuronal growth and survival linking it to neurodegenerative conditions like Alzheimer’s disease. Within these contexts RhoGDI interacts with other proteins such as Rac1 and Cdc42 to influence disease progression and pathology.
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5 product images
Western blot - Human ARHGDIA (RhoGDI) knockout HEK-293T cell lysate (ab257355)
Lane 1:Wild-type HEK293T cell lysate (20 ug)
Lane 2:ARHGDIA knockout HEK293T cell lysate (20 ug)
Lane 3:Jurkat cell lysate (20 ug)
Lane 4:HeLa cell lysate (20 ug)Anti-RhoGDI antibody [EPR3773] ab133248 was shown to specifically react with RhoGDI in wild-type HEK293T cells. Loss of signal was observed when knockout cell line Human ARHGDIA (RhoGDI) knockout HEK-293T cell line ab266446 (knockout cell lysate ab257355) was used. Wild-type and RhoGDI knockout samples were subjected to SDS-PAGE. Anti-RhoGDI antibody [EPR3773] ab133248 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated at room temperature for 2.5 hours at 1 in 1000 dilution 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-RhoGDI antibody [EPR3773] (Anti-RhoGDI antibody [EPR3773] ab133248) at 1/1000 dilution
Lane 1: Wild-type HEK293T cell lysate at 20 µg
Lane 2: ARHGDIA knockout HEK293T cell lysate at 20 µg
Lane 2: Western blot - Human ARHGDIA (RhoGDI) knockout HEK-293T cell line (Human ARHGDIA (RhoGDI) knockout HEK-293T cell line ab266446)
Lane 3: Jurkat cell lysate at 20 µg
Lane 4: HeLa cell lysate at 20 µg
SecondaryAll lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/10000 dilution
Predicted band size: 23 kDa
Observed band size: 23 kDa
Western blot - Human ARHGDIA (RhoGDI) knockout HEK-293T cell lysate (ab257355)
Lane 1:Wild-type HEK293T cell lysate (20 ug)
Lane 2:ARHGDIA knockout HEK293T cell lysate (20 ug)
Lane 3:Jurkat cell lysate (20 ug)
Lane 4:HeLa cell lysate (20 ug)Anti-RhoGDI antibody [EPR3772] ab108977 was shown to specifically react with RhoGDI in wild-type HEK293T cells. Loss of signal was observed when knockout cell line Human ARHGDIA (RhoGDI) knockout HEK-293T cell line ab266446 (knockout cell lysate ab257355) was used. Wild-type and RhoGDI knockout samples were subjected to SDS-PAGE. Anti-RhoGDI antibody [EPR3772] ab108977 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated at room temperature for 2.5 hours at 1 in 1000 dilution 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-RhoGDI antibody [EPR3772] (Anti-RhoGDI antibody [EPR3772] ab108977) at 1/1000 dilution
Lane 1: Wild-type HEK293T cell lysate at 20 µg
Lane 2: ARHGDIA knockout HEK293T cell lysate at 20 µg
Lane 2: Western blot - Human ARHGDIA (RhoGDI) knockout HEK-293T cell line (Human ARHGDIA (RhoGDI) knockout HEK-293T cell line ab266446)
Lane 3: Jurkat cell lysate at 20 µg
Lane 4: HeLa cell lysate at 20 µg
SecondaryAll lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/10000 dilution
Predicted band size: 21 kDa, 22 kDa, 23 kDa, 24 kDa, 25 kDa, 26 kDa, 28 kDa, 38 kDa, 46 kDa, 50 kDa, 57 kDa, 71 kDa
Observed band size: 22 kDa, 23 kDa, 25 kDa, 26 kDa, 39 kDa, 46 kDa, 71 kDa, 90 kDa
Western blot - Human ARHGDIA (RhoGDI) knockout HEK-293T cell lysate (ab257355)
Western blot: Anti-IRGM antibody (Anti-IRGM antibody ab69495) staining at 2 ug/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 magenta. In Western blot, Anti-IRGM antibody ab69495 was shown to bind specifically to IRGM. A band was observed at 18 kDa in wild-type A549 cell lysates with no signal observed at this size in IRGM knockout cell line. To generate this image, wild-type and IRGM knockout A549 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 °C. 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 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-RhoGDI antibody [2G3] (Anti-RhoGDI antibody [2G3] ab135252) at 1/1000 dilution
Lane 1: Wild-type HEK-293T cell lysate at 20 µg
Lane 2: ARHGDIA knockout HEK-293T cell lysate at 20 µg
Lane 2: Western blot - Human ARHGDIA (RhoGDI) knockout HEK-293T cell line (Human ARHGDIA (RhoGDI) knockout HEK-293T cell line ab266446)
Lane 2: Western blot - Human ARHGDIA (RhoGDI) knockout HEK-293T cell lysate (ab257355)
Lane 3: Wild-type HEK-293T ab255553 cell lysate at 20 µg
Lane 4: ARHGDIA knockout HEK-293T ab263544 cell lysate at 20 µg
Lane 5: HeLa cell lysate at 20 µg
Lane 6: U-87 MG cell lysate at 20 µg
Lane 7: Human Stomach cell lysate at 20 µg
Lane 8: Human Skeletal Muscle cell lysate at 20 µg
SecondaryAll lanes: Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Observed band size: 27 kDa
Sanger Sequencing - Human ARHGDIA (RhoGDI) knockout HEK-293T cell lysate (ab257355)
Allele-1: 1 bp insertion in exon 2
Sanger Sequencing - Human ARHGDIA (RhoGDI) knockout HEK-293T cell lysate (ab257355)
Allele-2: Insertion of the selection cassette in exon 2
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