Product nameHuman ARHGEF2 (GEF H1) knockout HeLa cell lysate
Access thousands of knockout cell lysates, generated from commonly used cancer cell lines.
See here for more information on knockout cell lysates.
User storage instructions: After reconstitution, store the lysate at -80°C.
Parental Cell LineHeLa
Mutation descriptionKnockout achieved by using CRISPR/Cas9, 1 bp insertion in exon4.
Knockout validationSanger Sequencing, Western Blot (WB)
Reconstitution notesTo use as WB control, resuspend the lyophilizate in 50 µL of LDS* Sample Buffer to have a final concentration of 2 mg/ml. For reducing conditions, we recommend a final concentration of 0.1 M DTT.
*Usage of SDS sample buffer is not recommended with these lyophilized lysates.
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.
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.
Tested applicationsSuitable for: WBmore details
Storage instructionsStore at -80°C. Please refer to protocols.
Components 1 kit Human ARHGEF2 knockout HeLa cell lysate (Lyophilized) 1 x 100µg Human Wild Type HeLa cell lysate (Lyophilized) 1 x 100µg
STR AnalysisAmelogenin X D5S818: 11, 12 D13S317: 12, 13.3 D7S820: 8, 12 D16S539: 9, 10 vWA: 16, 18 TH01: 7 TPOX: 8,12 CSF1PO: 9, 10
FunctionActivates Rho-GTPases by promoting the exchange of GDP for GTP. May be involved in epithelial barrier permeability, cell motility and polarization, dendritic spine morphology, antigen presentation, leukemic cell differentiation, cell cycle regulation, and cancer. Binds Rac-GTPases, but does not seem to promote nucleotide exchange activity toward Rac-GTPases, which was uniquely reported in PubMed:9857026. May stimulate instead the cortical activity of Rac. Inactive toward CDC42, TC10, or Ras-GTPases. Forms an intracellular sensing system along with NOD1 for the detection of microbial effectors during cell invasion by pathogens. Required for RHOA and RIP2 dependent NF-kappaB signaling pathways activation upon S.flexneri cell invasion. Involved not only in sensing peptidoglycan (PGN)-derived muropeptides through NOD1 that is independent of its GEF activity, but also in the activation of NF-kappaB by Shigella effector proteins (IpgB2 and OspB) which requires its GEF activity and the activation of RhoA.
Sequence similaritiesContains 1 DH (DBL-homology) domain.
Contains 1 PH domain.
Contains 1 phorbol-ester/DAG-type zinc finger.
DomainThe DH (DBL-homology) domain interacts with and promotes loading of GTP on RhoA.
The PH (pleckstrin-homology) domain is involved in microtubule binding and targeting to tight junctions.
modificationsPhosphorylation of Ser-886 by PAK1 induces binding to protein 14-3-3 zeta, promoting its relocation to microtubules and the inhibition of its activity. Phosphorylated by STK6 and CDK1 during mitosis, which negatively regulates its activity. Phosphorylation by MAPK1 or MAPK3 increases nucleotide exchange activity. Phosphorylation by PAK4 releases GEF-H1 from the microtubules.
Cellular localizationCytoplasm. Cell junction > tight junction. Golgi apparatus. Cytoplasm > cytoskeleton > spindle. Cell projection > ruffle membrane. Localizes to the tips of cortical microtubules of the mitotic spindle during cell division, and is further released upon microtubule depolymerization. Recruited into membrane ruffles induced by S.flexneri at tight junctions of polarized epithelial cells.
- Information by UniProt
KO cell lines
Our Abpromise guarantee covers the use of ab257223 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||Use at an assay dependent concentration. Predicted molecular weight: 111 kDa.|
Lane 1: Wild-type HeLa cell lysate (20 µg)
Lane 2: ARHGEF2 knockout HeLa cell lysate (20 µg)
Lane 3: HEK293 cell lysate (20 µg)
ab201687 Anti-GEF H1 antibody [EPR17963] - C-terminal was shown to specifically react with GEF H1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265797 (knockout cell lysate ab257223) was used. Wild-type and GEF H1 knockout samples were subjected to SDS-PAGE. ab201687 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C 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 10000 dilution for 1 hour at room temperature before imaging.
Homozygous: 1 bp insertion in exon4
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
ab257223 has not yet been referenced specifically in any publications.