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AB257637

Human RHOA knockout HEK-293T cell lysate

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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.
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Western blot - Human RHOA knockout HEK-293T cell lysate (AB257637)
  • WB

Lab

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 - ab187027 observed at 21 kDa. Red - loading control ab7291 observed at 50 kDa.

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 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. ab187027 and 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 (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-RhoA antibody [EPR18134] (<a href='/en-us/products/primary-antibodies/rhoa-antibody-epr18134-ab187027'>ab187027</a>) 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

Lane 2:

Western blot - Human RHOA knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-rhoa-knockout-hek-293t-cell-line-ab266592'>ab266592</a>)

Predicted band size: 22 kDa

Observed band size: 21 kDa

false

Sanger Sequencing - Human RHOA knockout HEK-293T cell lysate (AB257637)
  • Sanger seq

Unknown

Sanger Sequencing - Human RHOA knockout HEK-293T cell lysate (AB257637)

Allele-2 : Insertion of the selection cassette in exon 2

Sanger Sequencing - Human RHOA knockout HEK-293T cell lysate (AB257637)
  • Sanger seq

Unknown

Sanger Sequencing - Human RHOA knockout HEK-293T cell lysate (AB257637)

Allele-1 : 4 bp insertion 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.

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Complementary Products

Primary Antibodies

AB187027

Anti-RhoA antibody [EPR18134]

RabMAb|Recombinant|KO Validated

Western blot - Anti-RhoA antibody [EPR18134] (AB187027)

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Cell Lines & Lysates

AB266592

Human RHOA knockout HEK-293T cell line

Advanced Validation

Western blot - Human RHOA knockout HEK-293T cell line (AB266592)

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  • 0
Cell Lines & Lysates

AB150035

HeLa whole cell lysate

Western blot - HeLa whole cell lysate (AB150035)

  • 0
  • 0
Cell Lines & Lysates

AB14652

HeLa nuclear extract lysate (TNF alpha stimulated)

Functional Studies - HeLa nuclear extract lysate (TNF alpha stimulated) (AB14652)

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  • 0
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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

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?

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Properties and storage information

Gene name
RHOA
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.

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.

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.
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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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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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