Invitrogen

RhoGDI Monoclonal Antibody (2G3)

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

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Datasheet

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Datasheet

Questions & Answers

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RhoGDI Antibody in Immunofluorescence (IF) — Immunofluorescence analysis of HepG2 cells using ARHGDIA monoclonal antibody (Product # MA5-17032) (Green). Blue: DRAQ5 fluorescent DNA dye.

RhoGDI Antibody in Immunocytochemistry (ICC) — 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 (2G3) (Product # MA5-17032) 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 Cytoplasmic 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 Western Blot (WB) — Western blot was performed using Anti-RhoGDI Monoclonal Antibody (2G3)(Product # MA5-17032) and a 25kDa band corresponding to RhoGDI was observed across the cell lines tested. Whole Cell Extract-WCL (30 ug µg lysate) of SH-SY5Y (Lane 1), U-937 (Lane 2), NIH:OVCAR-3 (Lane 3), COS-7 (Lane 4), NIH/3T3 (Lane 5), HeLa (Lane 6) were electrophoresed using NuPAGE™ 12% Bis-Tris Protein Gel (Product # NP0342BOX). 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 in Flow Cytometry (Flow) — Flow cytometric analysis of HeLa cells using ARHGDIA monoclonal antibody (Product # MA5-17032) (green) and negative control (red).

RhoGDI Antibody in Knockdown — Antibody specificity was demonstrated by siRNA mediated KD 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 (2G3) (Product # MA5-17032). {KD}

Product Details

Applications

Tested Dilution

Publications

Flow Cytometry (Flow)

1:200 - 1:400

Immunocytochemistry (ICC)

1:100-1:1,000

Immunofluorescence (IF)

1:100-1:1,000

Western Blot (WB)

1:500 - 1:2,000

Product Specifications

Species

Human, Mouse, Non-human primate, Rat

Expression System

Mouse / IgG1

Class

Monoclonal

Type

Antibody

Clone

2G3

Immunogen

Purified recombinant fragment of human ARHGDIA (amino acids: FULL(1-204)) expressed in E. Coli.

Conjugate

Unconjugated

Form

Liquid

Concentration

1 mg/mL

Purification

purified

Storage buffer

PBS

Contains

0.05% sodium azide

Storage conditions

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

RRID

AB_2538504

Product Specific Information

MA5-17032 targets ARHGDIA in FACS, ICC, IF and WB applications and shows reactivity with Human, Mouse, and Non-human primate samples.

The MA5-17032 immunogen is purified recombinant fragment of human ARHGDIA (amino acids: FULL(1-204)) expressed in E. Coli.

MA5-17032 detects ARHGDIA which has a predicted molecular weight of approximately 26kDa.

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: arhgdia {ECO:0000312|RGD:1359547}; epididymis secretory sperm binding protein Li 47e; GDI-1; GDP-dissociation inhibitor, aplysia RAS-related 1; MGC117248; Rho GDI 1; rho GDI 1 {ECO:0000250|UniProtKB:P19803}; Rho GDI alpha; Rho GDP dissociation inhibitor (GDI) alpha; Rho GDP-dissociation inhibitor 1; rho GDP-dissociation inhibitor 1 {ECO:0000250|UniProtKB:P19803}; Rho-GDI alpha; rho-GDI alpha {ECO:0000250|UniProtKB:P19803}

Gene Aliases: 5330430M07Rik; ARHGDIA; Bles01; C87222; Gdi-1; Gdi1; GDIA1; HEL-S-47e; NPHS8; Rho-GDI; RhoDGI; RHOGDI; RHOGDI-1

UniProt ID: (Human) P52565, (Rat) Q5XI73, (Mouse) Q99PT1

Entrez Gene ID: (Human) 396, (Rat) 360678, (Mouse) 192662

Molecular Function: G-protein modulator enzyme modulator signaling molecule