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AB266447

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

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ARHGDIA KO cell line available to order. KO validated by Western blot. Free of charge wild type control provided. Knockout achieved by using CRISPR/Cas9, 1 bp deletion 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

4 Images
Western blot - Human ARHGDIA (RhoGDI) knockout HEK-293T cell line (AB266447)
  • WB

Lab

Western blot - Human ARHGDIA (RhoGDI) knockout HEK-293T cell line (AB266447)

Lanes 1-3 : Merged signal (red and green). Green - ab108977 observed at 27 kDa. Red - loading control ab8245 observed at 36 kDa.

ab108977 Anti-RhoGDI antibody [EPR3772] was shown to specifically react with RhoGDI in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266447 (knockout cell lysate ab257356) was used. Wild-type and RhoGDI knockout samples were subjected to SDS-PAGE. ab108977 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° 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 (ab266447)

Lane 3:

Jurkat 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,27 kDa,30 kDa,62 kDa

Observed band size: 27 kDa,30 kDa,70 kDa

false

Western blot - Human ARHGDIA (RhoGDI) knockout HEK-293T cell line (AB266447)
  • WB

Lab

Western blot - Human ARHGDIA (RhoGDI) knockout HEK-293T cell line (AB266447)

Lanes 1-4 : Merged signal (red and green). Green - ab133248 observed at 27 kDa. Red - loading control ab8245 observed at 36 kDa.

ab133248 Anti-RhoGDI antibody [EPR3773] was shown to specifically react with RhoGDI in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266447 (knockout cell lysate ab257356) was used. Wild-type and RhoGDI knockout samples were subjected to SDS-PAGE. ab133248 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4° 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 (ab266447)

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: 27 kDa

false

Sanger Sequencing - Human ARHGDIA (RhoGDI) knockout HEK-293T cell line (AB266447)
  • Sanger seq

Unknown

Sanger Sequencing - Human ARHGDIA (RhoGDI) knockout HEK-293T cell line (AB266447)

Allele-2 : 1 bp deletion in exon 2.

Sanger Sequencing - Human ARHGDIA (RhoGDI) knockout HEK-293T cell line (AB266447)
  • Sanger seq

Unknown

Sanger Sequencing - Human ARHGDIA (RhoGDI) knockout HEK-293T cell line (AB266447)

Allele-1 : Insertion of the selection cassette in exon2

Key facts

Cell type

HEK-293T

Species or organism

Human

Tissue

Kidney

Form

Liquid

form

Knockout validation

Sanger Sequencing,Western blot

Mutation description

Knockout achieved by using CRISPR/Cas9, 1 bp deletion in exon 2 and Insertion of the selection cassette in exon 2

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "WB": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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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.

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What's included?

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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
Dry Ice
Appropriate short-term storage conditions
-196°C
Appropriate long-term storage conditions
-196°C
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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.

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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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Target data

See full target information ARHGDIA
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