PA5-21429

antibody from Invitrogen Antibodies

Targeting: ARHGEF18 KIAA0521, MGC15913, P114-RhoGEF, p114RhoGEF

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-21429

Provider

Invitrogen Antibodies

Product name

ARHGEF18 Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant protein fragment

Description

Recommended positive controls: HepG2.

Concentration

1.0 mg/mL

References

Submitted references

GPCR-independent activation of G proteins promotes apical cell constriction in vivo.

Marivin A, Morozova V, Walawalkar I, Leyme A, Kretov DA, Cifuentes D, Dominguez I, Garcia-Marcos M
The Journal of cell biology 2019 May 6;218(5):1743-1763

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of ARHGEF18 using 30 µg of HepG2 lysate. Samples were loaded onto a 7.5% SDS-PAGE gel and probed with an ARHGEF18 polyclonal antibody (Product # PA5-21429) at a dilution of 1:1000.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot using ARHGEF18 Polyclonal Antibody (Product # PA5-21429). Various whole cell extracts (30 µg) were separated by 5% SDS-PAGE, and the membrane was blotted with ARHGEF18 Polyclonal Antibody (Product # PA5-21429) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot using ARHGEF18 Polyclonal Antibody (Product # PA5-21429). Whole cell extract (30 µg) was separated by 5% SDS-PAGE, and the membrane was blotted with ARHGEF18 Polyclonal Antibody (Product # PA5-21429) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent analysis of ARHGEF18 in methanol-fixed A431 cells using an ARHGEF18 polyclonal antibody (Product # PA5-21429) at a 1:200 dilution.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of methanol-fixed A431, using ARHGEF18 antibody (Product # PA5-21429) at 1:200 dilution.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of methanol-fixed A431, using ARHGEF18 antibody (Product # PA5-21429) at 1:200 dilution.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry (Paraffin) analysis of ARHGEF18 was performed in paraffin-embedded mouse thymus gland tissue using ARHGEF18 Polyclonal Antibody (Product # PA5-21429) at a dilution of 1:500.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemistry (Paraffin) analysis of ARHGEF18 was performed in paraffin-embedded rat spleen tissue using ARHGEF18 Polyclonal Antibody (Product # PA5-21429) at a dilution of 1:500.

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Both the PBM and GBA motif of DAPLE are required to promote apical cell constriction. (A) Quantification of the relative apical area of DAPLE-transfected cells compared with neighboring, untransfected cells shows that hDAPLE ΔPBM and hDAPLE GBA* mutants fail to promote apical constriction in MDCK (left) or EpH4 (right) cells. Results are presented as box-and-whiskers plots (error bars indicate minimum to maximum range) of n = 4-9 independent experiments per condition. *, P < 0.05; **, P < 0.01 using the Mann-Whitney U test. (B) Confocal fluorescence microscopy pictures of MDCK cells sparsely expressing the indicated MYC-hDAPLE constructs and costained for MYC (magenta) and ZO-1 (green) show that hDAPLE GBA* mutant is enriched at apical junctions like WT, while hDAPLE ΔPBM mutant is not. Scale bars, 10 µm. (C) Diagram depicting the different functions of the PBM and the GBA motif in DAPLE-induced apical cell constriction. A possible effector pathway to promote apical cell constriction though G protein activation is shown on the right, along with the treatments used in D to test it (green). (D) Box-and-whiskers plots (error bars indicate minimum to maximum range) for the quantification of relative apical area show that DAPLE-mediated apical constriction requires the activity of myosin (inhibited by blebbistatin), ROCK (inhibited by Y-27632), free Gßγ (inhibited by Gallein, but not its inactive analogue, fluorescein) and p114RhoGEF (inhibited by siRNA). n = 4 independent experim