IQGAP1 antibody

Catalog No. ABIN487492

This Mouse Monoclonal antibody specifically detects IQGAP1 in WB and IP. It exhibits reactivity toward Human, Mouse and Rat.

Quick Overview for IQGAP1 antibody (ABIN487492)

Target: IQGAP1 (IQ Motif Containing GTPase Activating Protein 1 (IQGAP1))

Reactivity: Human, Mouse, Rat

Host: Mouse

Clonality: Monoclonal

Conjugate: This IQGAP1 antibody is un-conjugated

Application: Western Blotting (WB), Immunoprecipitation (IP)

Clone: VIC1-IC11-IVB2

Product Details anti-IQGAP1 Antibody

Specificity: This antibody reacts with Human, Mouse and Rat IQGAP1.

Cross-Reactivity (Details): Species reactivity (tested):Human, Mouse and Rat.

Characteristics: Synonyms: KIAA0051, p195, Ras GTPase-activating-like protein IQGAP1

Purification: Protein-A Agarose Chromatography of hybridoma supernatant.

Immunogen: Purified native Bovine adrenal IQGAP1. Remarks: Hybridoma was established by fusion of mouse plasmacytoma ell NS-1 withBalb/c mouse lymph nodes.

Isotype: IgG2a

Application Details

Application Notes: Western blot: 1 μg/mL for chemiluminescence detection system. Positive Controls: HeLa, A431, NIH/3T3, C2C12, Rat-1. Immunoprecipitation: 2 μg/200-300 μL of cell extract. Positive Control: HeLa. Detailed procedure is provided in Protocols.
Other applications not tested.
Optimal dilutions are dependent on conditions and should be determined by the user.

Protocol: SDS-PAGE & Western Blotting1) Wash the cells 3 times with PBS and suspend with 10 volume of cold Lysis buffer (50 mMTris-HCl, pH 7. 2, 250 mM NaCl, 0. 1% NP-40, 2 mM EDTA, 10% glycerol) containingappropriate protease inhibitors. Incubate it at 4°C with rotating for 30 minutes, thensonicate briefly (up to 10 seconds). 2) Centrifuge the tube at 12,000 x g for 10 minutes at 4°C and transfer the supernatant toanother tube. Measure the protein concentration of the supernatant and add the Lysisbuffer to make 8 mg/mL solution. 3) Mix the sample with equal volume of Laemmli’s sample buffer. 4) Boil the samples for 2 minutes and centrifuge. Load 10 μL of the sample per lane in a 1mm thick SDS-polyacrylamide gel for electrophoresis. 5) Blot the protein to a polyvinylidene difluoride (PVDF) membrane at 1 mA/cm2 for 1 hourin a semi-dry transfer system. (Transfer Buffer: 25 mM Tris, 190 mM glycine, 20% MeOH). See the manufacture's manual for the transfer procedure. 6) To reduce nonspecific binding, soak the membrane in 10% skimmed milk (in PBS, pH7. 2) for 1 hour at room temperature, or overnight at 4°C. 7) Incubate the membrane with primary antibody diluted with PBS, pH 7. 2 containing 1%skimmed milk as suggested in the APPLICATIONS for 1 hour at room temperature. (Theoptimal antibody concentration will depend on the experimental conditions. )8) Wash the membrane with PBS (5 minutes x 6 times). 9) Incubate the membrane with the 1: 10000 POD-conjugated anti-mouse IgG diluted with1% skimmed milk (in PBS, pH 7. 2) for 1 hour at room temperature. 10) Wash the membrane with PBS (5 minutes x 6 times). 11) Wipe excess buffer from the membrane, then incubate it with appropriatechemiluminescence reagents for 1 minute. Remove extra reagent from the membrane bydabbing with a paper towel, and seal it in plastic wrap. 12) Expose to an X-ray film in a dark room for 5 minutes. Develop the film as usual. Theconditions for exposure and development may vary. Positive Controls for Western blotting: HeLa, A431, NIH/3T3, C2C12, Rat-1. Immunoprecipitation: 1) Wash the cells 3 times with PBS and suspend with 10 volume of cold Lysis buffer (50 mMTris-HCl, pH 7. 2, 250 mM NaCl, 0. 1% NP-40, 2 mM EDTA, 10% glycerol) containingappropriate protease inhibitors. Incubate it at 4°C with rotating for 30 minutes, thensonicate briefly (up to 10 seconds). 2) Centrifuge the tube at 12,000 x g for 10 minutes at 4°C and transfer the supernatant toanother tube. 3) Add 2 µg of the anti-IQGAP1 monoclonal antibody into 250 µL of the supernatant. Mixwell and incubate with gentle agitation for 30-120 minutes at 4°C. 4) Add 20 µL of 50% protein A agarose beads resuspended in the Lysis buffer. Mix well andincubate with gentle agitation for 60 minutes at 4°C. 5) Wash the beads 3-5 times with ice-cold Lysis buffer (centrifuge the tube at 2,500 x g for10 seconds). 6) Resuspend the beads in 20 µL of Laemmli’s sample buffer, boil for 3-5 minutes, andcentrifuge for 5 minutes. Use 10 µL/lane for the SDS-PAGE analysis. (See SDS-PAGE & Western blotting. )Positive Control for Immunoprecipitation: HeLa.

Restrictions: For Research Use only

Handling

Concentration: 1.0 mg/mL

Buffer: PBS, pH 7.2 containing 50 % Glycerol without preservatives.

Preservative: Without preservative

Storage: -20 °C

Storage Comment: Store the antibody (in aliquots) at -20 °C. Avoid repeated freezing and thawing.
Shelf life: one year from despatch.

Expiry Date: 12 months

Target Details for IQGAP1

Target: IQGAP1 (IQ Motif Containing GTPase Activating Protein 1 (IQGAP1))

Alternative Name: IQGAP1

Background: IQ domain GTPase activating protein (IQGAP) is a ~170 kDa protein that is a target molecule for activated Cdc42 and Rac1. IQGAP binds actin and calmodulin and is an effector for the Rho family of GTPases, providing a direct link between the activated GTPase and the actin cytoskeleton. Additionally, the calmodulin-binding protein IQGAP1 binds stoichiometrically to beta-catenin, an oncoprotein integral to cell-cell adhesion and proliferative signaling, and regulates the association of beta-catenin with the cell-cell adhesion complex.Synonyms: KIAA0051, Ras GTPase-activating-like protein IQGAP1, p195

Gene ID: 8826

UniProt: P46940

Pathways: Signaling Events mediated by VEGFR1 and VEGFR2