Invitrogen

RhoGDI Monoclonal Antibody (OTI1A7)

Advanced Verification

This Antibody was verified by Knockdown to ensure that the antibody binds to the antigen stated.

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Datasheet

Protocols

Questions & Answers

Tech Support

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RhoGDI Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of RhoGDI was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with RhoGDI Monoclonal Antibody (OTI1A7) (Product # MA5-25090) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32766), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300 dilution). Panel d represents the merged image showing Nuclear and cytosolic localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X with Oil immersion magnification.

RhoGDI Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry was performed on paraffin-embedded adenocarcinoma of colon tissue. To expose target proteins, 10mM citric buffer, pH6.0, 100°C for 10min was used. Following antigen retrieval, tissues were probed with a ARHGDIA monoclonal antibody (Product # MA5-25090) at a dilution of 1:50.

RhoGDI Antibody in Western Blot (WB) — Western blot was performed using Anti-RhoGDI Monoclonal Antibody (OTI1A7)(Product # MA5-25090) and a 25 kDa band corresponding to RhoGDI was observed across the cell lines tested. Whole cell extracts (30 µg lysate) of HEK-293 (Lane 1), SH-SY5Y (Lane 2), U-937 (Lane 3), NIH:OVCAR-3 (Lane 4), COS-7 (Lane 5), NIH/3T3 (Lane 6) and HeLa (Lane 7) were electrophoresed using NuPAGE™ 12% Bis-Tris Protein Gel (Product # NP0341BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # LC2001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177,1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

RhoGDI Antibody — Antibody specificity was demonstrated by siRNA mediated knockdown of target protein. HeLa cells were transfected with RhoGDI siRNA and decrease in signal intensity was observed in Western Blot application using Anti-RhoGDI Monoclonal Antibody (OTI1A7) (Product # MA5-25090). {KD}

Product Details

MA5-25090

Applications	Tested Dilution	Publications
Western Blot (WB)	1:1,000-1:2,000	-
Immunohistochemistry (Paraffin) (IHC (P))	1:50	-
Immunocytochemistry (ICC/IF)	1:100	-

Product Specifications
Species Reactivity	Human
Host/Isotype	Mouse / IgG1
Class	Monoclonal
Type	Antibody
Clone	OTI1A7
Immunogen	Full length human recombinant protein of ARHGDIA produced in HEK293T cell View antigen
Conjugate	Unconjugated
Form	Liquid
Concentration	0.72 mg/mL
Purification	Affinity Chromatography
Storage buffer	PBS, pH 7.3, with 1% BSA, 50% glycerol
Contains	0.02% sodium azide
Storage conditions	-20° C, Avoid Freeze/Thaw Cycles
RRID	AB_2725231

Target Information

Aplysia Ras-related homologs (ARHs), also called Rho genes, belong to the RAS gene superfamily encoding small guanine nucleotide exchange (GTP/GDP) factors. The ARH proteins may be kept in the inactive, GDP-bound state by interaction with GDP dissociation inhibitors, such as ARHGDIA.

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

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Bioinformatics

Protein Aliases: epididymis secretory sperm binding protein Li 47e; GDP-dissociation inhibitor, aplysia RAS-related 1; MGC117248; Rho GDI 1; Rho GDP dissociation inhibitor (GDI) alpha; Rho GDP-dissociation inhibitor 1; Rho-GDI alpha

Gene Aliases: ARHGDIA; GDIA1; HEL-S-47e; NPHS8; RHOGDI; RHOGDI-1

UniProt ID: (Human) P52565

Entrez Gene ID: (Human) 396

Molecular Function: G-protein modulator