Invitrogen

IQGAP1 Polyclonal Antibody

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IQGAP1 Antibody in Immunocytochemistry (ICC/IF) — Immunocytochemistry analysis of IQGAP1 using anti-IQGAP1 antibody (Product # PA5-95625) . IQGAP1 was detected in a section of U2OS cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum and then incubated with 2μg/mL rabbit anti-IQGAP1 antibody (Product # PA5-95625) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

IQGAP1 Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry analysis of IQGAP1 in paraffin-embedded human intestinal cancer tissue. Antigen retrieval was performed on the tissue using citrate buffer (pH 6, 20 min) and blocked with 10% goat serum. Samples were incubated with IQGAP1 polyclonal antibody (Product # PA5-95625) at a 1 µg/mL dilution, followed by biotinylated goat anti-rabbit IgG (30 min, 37°C), and developed with Strepavidin-Biotin-Complex and DAB.

IQGAP1 Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry analysis of IQGAP1 in paraffin-embedded human intestinal cancer tissue. Antigen retrieval was performed on the tissue using citrate buffer (pH 6, 20 min) and blocked with 10% goat serum. Samples were incubated with IQGAP1 polyclonal antibody (Product # PA5-95625) at a 1 µg/mL dilution, followed by biotinylated goat anti-rabbit IgG (30 min, 37°C), and developed with Strepavidin-Biotin-Complex and DAB.

IQGAP1 Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry analysis of IQGAP1 in paraffin-embedded human mammary cancer tissue. Antigen retrieval was performed on the tissue using citrate buffer (pH 6, 20 min) and blocked with 10% goat serum. Samples were incubated with IQGAP1 polyclonal antibody (Product # PA5-95625) at a 1 µg/mL dilution, followed by biotinylated goat anti-rabbit IgG (30 min, 37°C), and developed with Strepavidin-Biotin-Complex and DAB.

IQGAP1 Antibody in Immunohistochemistry (Paraffin) (IHC (P)) — Immunohistochemistry analysis of IQGAP1 in paraffin-embedded human mammary cancer tissue. Antigen retrieval was performed on the tissue using citrate buffer (pH 6, 20 min) and blocked with 10% goat serum. Samples were incubated with IQGAP1 polyclonal antibody (Product # PA5-95625) at a 1 µg/mL dilution, followed by biotinylated goat anti-rabbit IgG (30 min, 37°C), and developed with Strepavidin-Biotin-Complex and DAB.

IQGAP1 Antibody in Western Blot (WB) — Western blot was performed using Anti-IQGAP1 Polyclonal Antibody (Product # PA5-95625) and a 189 kDa band corresponding to IQGAP1 was observed across all cell lysates tested. Whole cell extracts (60 µg lysate) of HaCaT (Lane 1), THP-1 (Lane 2), K-562 (Lane 3), HEK-293 (Lane 4), A-431 (Lane 5), NIH/3T3 (Lane 6), Hep G2 (Lane 7) were electrophoresed using NuPAGE™ 3-8% Tris-Acetate Protein Gel (Product # EA0378BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1,000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036,1:10,000 dilution) using the iBright™ FL1500 Imaging System (Product # A44115). Chemiluminescent detection was performed using SuperSignal™ West Pico PLUS Chemiluminescent Substrate (Product # 34580). Uncharacterized bands (*) were observed at ~268 kDa, ~160 kDa and 80 kDa in some cell lines.

IQGAP1 Antibody in Western Blot (WB) — Knockdown of IQGAP1 was achieved by transfecting HEK-293 with IQGAP1 specific siRNAs (Silencer® select Product # s16837, s16839). Western blot analysis (Fig. a) was performed using Whole cell extracts from the IQGAP1 knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with IQGAP1 Polyclonal Antibody (Product # PA5-95625, 1:1,000 dilution) and Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:10,000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to IQGAP1. An uncharacterized band was observed at ~55 kDa in all cell lysates, while a faint uncharacterized band was observed at ~45 kDa in untransfected and scrambled siRNA lysates.

IQGAP1 Antibody in Western Blot (WB) — Western blot analysis of IQGAP1 in Lane 1: human placenta tissue lysates, Lane 2: human U-87MG whole cell lysates. Electrophoresis was performed with 5-20% SDS-PAGE gel (70V, Stacking gel; 90V Resolving gel, Time: 2-3 hours), transferred to a nitrocellulose membrane and blocked using 5% Non-fat Milk/TBS (1.5 hrs at room temperature). Samples were incubated with IQGAP1 polyclonal antibody (Product # PA5-95625) using a 0.5 µg/mL dilution, followed by a goat anti-rabbit IgG-HRP at a dilution of 1:10,000, and developed with enhanced chemiluminescence (ECL).

IQGAP1 Antibody in Flow Cytometry (Flow) — Flow Cytometry of IQGAP1 in HepG2 cells (blue line), isotype control rabbit IgG (green line) and unlabeled (red line). Samples were blocked with 10% goat serum, incubated with IQGAP1 Polyclonal Antibody (Product # PA5-95625) at a dilution of 1 μg (per 1x10^6 cells), followed by DyLight®488 conjugated goat anti-rabbit IgG (for 30 minutes at 20°C) using 5-10 μg (per 1x10^6 cells) dilution.

Product Details

PA5-95625

Applications	Tested Dilution	Publications
Western Blot (WB)	0.1-0.5 µg/mL	-
Immunohistochemistry (Paraffin) (IHC (P))	0.5-1 µg/mL	-
Immunocytochemistry (ICC/IF)	2 µg/mL	-
Flow Cytometry (Flow)	1-3 µg/1x10^6 cells	-
ELISA (ELISA)	0.1-0.5 µg/mL	-

Product Specifications
Species Reactivity	Human, Mouse, Rat
Host/Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	E.coli-derived human IQGAP1 recombinant protein (D1374-K1657). View immunogen
Conjugate	Unconjugated
Form	Lyophilized
Concentration	500 µg/mL
Purification	Affinity chromatography
Storage buffer	PBS with 4mg trehalose
Contains	0.05mg sodium azide
Storage conditions	Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
RRID	AB_2807427

Product Specific Information

Reconstitute with 0.2 mL of distilled water to yield a concentration of 500 µg/mL.

Target Information

This gene encodes a member of the IQGAP family. The protein contains four IQ domains, one calponin homology domain, one Ras-GAP domain and one WW domain. It interacts with components of the cytoskeleton, with cell adhesion molecules, and with several signaling molecules to regulate cell morphology and motility. Expression of the protein is upregulated by gene amplification in two gastric cancer cell lines.

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

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Bioinformatics

Protein Aliases: Cdc42-Rac1 effector protein; DISP2; DISPB; p195; Ras GTPase-activating-like protein IQGAP1; RasGAP-like with IQ motifs

Gene Aliases: AA682088; D7Ertd237e; D7Ertd257e; HUMORFA01; IQGAP1; KIAA0051; mKIAA0051; p195; SAR1

UniProt ID: (Human) P46940, (Mouse) Q9JKF1

Entrez Gene ID: (Human) 8826, (Mouse) 29875, (Rat) 361598

Molecular Function: G-protein modulator protein-binding activity modulator GTPase-activating protein

regulation of cytokine production signal transduction epidermal growth factor receptor signaling pathway fibroblast growth factor receptor signaling pathway negative regulation of GTPase activity negative regulation of dephosphorylation cellular response to platelet-derived growth factor stimulus positive regulation of GTPase activity positive regulation of protein kinase activity platelet-derived growth factor receptor signaling pathway cellular response to calcium ion cellular response to epidermal growth factor stimulus positive regulation of protein serine/threonine kinase activity glomerular visceral epithelial cell development cell-cell adhesion neuron projection extension small GTPase mediated signal transduction regulation of GTPase activity cellular response to fibroblast growth factor stimulus

Process(es)

GTPase inhibitor activity GTPase activator activity calcium ion binding protein binding calmodulin binding phosphatidylinositol-3,4,5-trisphosphate binding protein kinase binding protein phosphatase binding protein domain specific binding protein complex binding protein serine/threonine kinase activator activity S100 protein binding Rac GTPase binding cadherin binding involved in cell-cell adhesion Rho GTPase binding