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AB257355

Human ARHGDIA (RhoGDI) knockout HEK-293T cell lysate

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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.

View 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

5 Images
Western blot - Human ARHGDIA (RhoGDI) knockout HEK-293T cell lysate (AB257355)
  • WB

Lab

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)

ab133248 was shown to specifically react with RhoGDI in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266446 (knockout cell lysate ab257355) was used. Wild-type and RhoGDI knockout samples were subjected to SDS-PAGE. ab133248 and 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 (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-RhoGDI antibody [EPR3773] (<a href='/en-us/products/primary-antibodies/rhogdi-antibody-epr3773-ab133248'>ab133248</a>) 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 (<a href='/en-us/products/cell-lines/human-arhgdia-rhogdi-knockout-hek-293t-cell-line-ab266446'>ab266446</a>)

Lane 3:

Jurkat cell lysate at 20 µg

Lane 4:

HeLa cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 23 kDa

Observed band size: 23 kDa

false

Western blot - Human ARHGDIA (RhoGDI) knockout HEK-293T cell lysate (AB257355)
  • WB

Lab

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)

ab108977 was shown to specifically react with RhoGDI in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266446 (knockout cell lysate ab257355) was used. Wild-type and RhoGDI knockout samples were subjected to SDS-PAGE. ab108977 and 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 (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-RhoGDI antibody [EPR3772] (<a href='/en-us/products/primary-antibodies/rhogdi-antibody-epr3772-ab108977'>ab108977</a>) 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 (<a href='/en-us/products/cell-lines/human-arhgdia-rhogdi-knockout-hek-293t-cell-line-ab266446'>ab266446</a>)

Lane 3:

Jurkat cell lysate at 20 µg

Lane 4:

HeLa cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) 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

false

Western blot - Human ARHGDIA (RhoGDI) knockout HEK-293T cell lysate (AB257355)
  • WB

Lab

Western blot - Human ARHGDIA (RhoGDI) knockout HEK-293T cell lysate (AB257355)

Western blot : Anti-IRGM antibody (ab69495) staining at 2 ug/ml, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, 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] (<a href='/en-us/products/primary-antibodies/rhogdi-antibody-2g3-ab135252'>ab135252</a>) 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 (<a href='/en-us/products/cell-lines/human-arhgdia-rhogdi-knockout-hek-293t-cell-line-ab266446'>ab266446</a>)

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

Secondary

All lanes:

Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution

Observed band size: 27 kDa

false

Sanger Sequencing - Human ARHGDIA (RhoGDI) knockout HEK-293T cell lysate (AB257355)
  • Sanger seq

Unknown

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)
  • Sanger seq

Unknown

Sanger Sequencing - Human ARHGDIA (RhoGDI) knockout HEK-293T cell lysate (AB257355)

Allele-2 : 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.

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Reactivity data

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Product details

Knockout cell lysate achieved by CRISPR/Cas9.

REACH authorisation
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.

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What's included?

{ "values": { "1Kit": { "sellingSize": "1 Kit", "publicAssetCode":"ab257355-1Kit", "assetComponentDetails": [ { "size":"1 x 100 µg", "name":"Human ARHGDIA knockout HEK293T cell lysate", "number":"AB257355-CMP01", "productcode":"" }, { "size":"1 x 100 µg", "name":"Human wild-type HEK293T cell lysate", "number":"AB257355-CMP02", "productcode":"" } ] } } }
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Properties and storage information

Gene name
ARHGDIA
Gene editing type
Knockout
Gene editing method
CRISPR technology
Knockout validation
Sanger Sequencing, Western blot
Shipped at conditions
Ambient - Can Ship with Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

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.

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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Quality control

STR analysis

CSF1PO, D13S317, D7S820, D5S818, TH01, D16S539, TPOX

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Cell culture

Biosafety level

EU: 2 US: 2

Adherent/suspension

Adherent

Gender

Female

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Product protocols

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