Invitrogen

NGEF Polyclonal Antibody

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NGEF Antibody in Immunohistochemistry (IHC) — Fig. 1 Ephexin1 is upregulated in CRC and LC and promotes tumor cell growth and proliferation. A Expression of Ephexin1 mRNA in 12 primary cancer types from TCGA. Data are shown as mean +- SD. ns not significant, *** P < 0.001, two-tailed Student's t test. B Kaplan-Meier analysis of overall survival according to Ephexin1 in CRC and LC patients. P values are for a log-rank test. C Immunohistochemistry (IHC) staining was performed to evaluate Ephexin1 expression in CRC and LC tissues and their corresponding normal tissues. Hematoxylin is the counterstain. Scale bar = 100 mum. Data are shown as mean +- SD, P values are for a two-way ANOVA. D Ephexin1 protein expression in normal and indicated colon ( n = 8) and lung ( n = 11) cancer cell lines were examined by western blot. E Effect of Ephexin1 knockdown on cell proliferation by MTT assay in HCT116, H1299, H460, and A549 cells. Data are shown as mean +- SD. *** P < 0.001, two-way ANOVA. F , G Anchorage-independent growth assessed by soft agar assay ( F ) and migration ability ( G ) determined by transwell assay in Ephexin1-depleted HCT116, H1299, H460, and A549 cells. Representative images are shown. Scale bar = 100 mum. Data are shown as mean +- SD. ** P < 0.01, and *** P < 0.001, two-tailed Student's t test. H Control and Ephexin1-depleted HCT116, H1299, and H460 cells were inoculated subcutaneously in BALB/C nude mice ( n = 4). Tumor volumes recoded at the indicated times are shown. Data are shown as mean +- SD. *** P < 0.001

NGEF Antibody in Western Blot (WB) — Fig. 2 Ephexin1 enhances the MEK/ERK signaling pathway and regulates K-Ras-downstream target genes. A Whole-cell lysates from control or Ephexin1-depleted H1299 cells were incubated with human phosphokinase array membranes. Each pair of the most positive kinase dots is numbered with the identification of the corresponding kinase listed. B Levels of indicated protein kinases were quantified after normalized positive control. Error bars indicate mean +- SD of spots. C Western blot analysis to detect MEK/ERK activation in indicated cells treated with EGF for the indicated amounts of time after serum starvation. D The co-expression heat map of Ephexin1 with K-Ras, H-Ras, N-Ras, or Ki-67 in TCGA and GTEx ( n = 5151) derived from the UCSC Xena browser. E Heat map showing the differential expressed genes induced by Ephexin1 shRNA in H1299 cells. Red and green indicate high and low mRNA expression levels, respectively. F Scatter plot shows the differently expressed genes (FDR <= 0.025 and fold difference >= 2) in Ephexin1-depleted H1299 cells compared with control cells. Compared to the control group (shControl), shEphexin1 downregulates 155 genes, whereas upregulates 227 genes expression levels. White circles highlight the position of the indicated K-Ras-downstream target genes. G Gene set enrichment analysis (GSEA) was used to identify K-Ras-downstream target genes differentially expressed between control and Ephexin1-depleted H1299 cells. H Heatmap of Ras-target genes downregulate

Product Details

PA5-52521

Applications	Tested Dilution	Publications
Western Blot (WB)	-	View 1 publication 1 publication
Immunohistochemistry (IHC)	1:50-1:200	View 2 publications 2 publications

Product Specifications
Species Reactivity	Human
Published species	Mouse
Host/Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	Recombinant protein corresponding to Human NGEF
Conjugate	Unconjugated
Form	Liquid
Concentration	0.1 mg/mL
Purification	Antigen affinity chromatography
Storage buffer	PBS, pH 7.2, with 40% glycerol
Contains	0.02% sodium azide
Storage conditions	Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
RRID	AB_2644657

Product Specific Information

Immunogen sequence: METRESEDLE KTRRKSASDQ WNTDNEPAKV KPELLPEKEE TSQADQDIQD KEPHCHIPIK RNSIFNRSIR RKSKAKARDN PERNASCLAD SQDNGKSVNE PLTLNIPWSR TPPCRTAMQT DPGAQEMSES SST

Highest antigen sequence identity to the following orthologs: Mouse - 48%, Rat - 26%.

Target Information

Acts as a guanine nucleotide exchange factor (GEF) which differentially activates the GTPases RHOA, RAC1 and CDC42. Plays a role in axon guidance regulating ephrin-induced growth cone collapse and dendritic spine morphogenesis. Upon activation by ephrin through EPHA4, the GEF activity switches toward RHOA resulting in its activation. Activated RHOA promotes cone retraction at the expense of RAC1- and CDC42-stimulated growth cone extension.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

Interactions between EGFR and EphA2 promote tumorigenesis through the action of Ephexin1.

Cell death & disease

Kim J,Chang IY,You HJ

PA5-52521 was used in Immunohistochemistry to show that the formation of a complex between EGFR, EphA2, and Ephexin1 plays an important role in lung and colorectal cancers, and that inhibition of this complex may be an effective target for cancer therapy.

Mon Jun 06 00:00:00 EDT 2022

Akt-mediated Ephexin1-Ras interaction promotes oncogenic Ras signaling and colorectal and lung cancer cell proliferation.

Cell death & disease

Kim J,Jeon YJ,Lim SC,Ryu J,Lee JH,Chang IY,You HJ

Published figure using NGEF polyclonal antibody (Product # PA5-52521) in Western Blot

Thu Oct 28 00:00:00 EDT 2021

Akt-mediated Ephexin1-Ras interaction promotes oncogenic Ras signaling and colorectal and lung cancer cell proliferation.

Cell death & disease

Kim J,Jeon YJ,Lim SC,Ryu J,Lee JH,Chang IY,You HJ

Published figure using NGEF polyclonal antibody (Product # PA5-52521) in Western Blot

Thu Oct 28 00:00:00 EDT 2021

Bioinformatics

Protein Aliases: Eph-interacting exchange protein; Ephexin-1; Neuronal guanine nucleotide exchange factor

Gene Aliases: ARHGEF27; EPHEXIN; NGEF

UniProt ID: (Human) Q8N5V2

Entrez Gene ID: (Human) 25791

Molecular Function: guanyl-nucleotide exchange factor

Function(s)

regulation of Rho protein signal transduction positive regulation of apoptotic process regulation of GTPase activity positive regulation of GTPase activity ephrin receptor signaling pathway regulation of small GTPase mediated signal transduction negative regulation of dendritic spine morphogenesis

Process(es)

guanyl-nucleotide exchange factor activity Rho guanyl-nucleotide exchange factor activity

NGEF Polyclonal Antibody

Product Details

PA5-52521

Western Blot (WB)

Immunohistochemistry (IHC)

Species Reactivity

Published species

Host/Isotype

Class

Type

Immunogen

Conjugate

Form

Concentration

Purification

Storage buffer

Contains

Storage conditions

RRID

Product Specific Information

Target Information

Bioinformatics

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