IQGAP1 抗体
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Quick Overview for IQGAP1 抗体 (ABIN487492)
抗原
See all IQGAP1 抗体适用
宿主
克隆类型
标记
应用范围
克隆位点
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特异性
- This antibody reacts with Human, Mouse and Rat IQGAP1.
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交叉反应 （详细）
- Species reactivity (tested):Human, Mouse and Rat.
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产品特性
- Synonyms: KIAA0051, p195, Ras GTPase-activating-like protein IQGAP1
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纯化方法
- Protein-A Agarose Chromatography of hybridoma supernatant.
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免疫原
- Purified native Bovine adrenal IQGAP1. Remarks: Hybridoma was established by fusion of mouse plasmacytoma ell NS-1 withBalb/c mouse lymph nodes.
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亚型
- IgG2a
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anti-IQ Motif Containing GTPase Activating Protein 1 (IQGAP1) (AA 500-750) antibody
IQGAP1 适用: 人 WB, IHC, IF 宿主: 兔 Polyclonal unconjugated
anti-IQ Motif Containing GTPase Activating Protein 1 (IQGAP1) antibodyKD Validated IQGAP1 适用: 人 WB, FACS 宿主: 兔 Monoclonal 25GB2050 unconjugated Recombinant Antibody
anti-IQ Motif Containing GTPase Activating Protein 1 (IQGAP1) (AA 611-710) antibodyIQGAP1 适用: 人 WB, ELISA, IF, IHC (p) 宿主: 小鼠 Monoclonal 2C5 unconjugated
anti-IQ Motif Containing GTPase Activating Protein 1 (IQGAP1) (AA 1451-1550) antibodyIQGAP1 适用: 人, 大鼠, 兔 WB, ELISA, IF (p), IF (cc), IHC (p), IHC (fro) 宿主: 兔 Polyclonal unconjugated
anti-IQ Motif Containing GTPase Activating Protein 1 (IQGAP1) antibodyIQGAP1 适用: 人, 小鼠, 大鼠 IHC, IF 宿主: 兔 Polyclonal unconjugated
anti-IQ Motif Containing GTPase Activating Protein 1 (IQGAP1) (N-Term) antibodyIQGAP1 适用: 人, 小鼠, 大鼠 WB, ELISA, IF, ICC 宿主: 兔 Polyclonal unconjugated
anti-IQ Motif Containing GTPase Activating Protein 1 (IQGAP1) antibodyKD Validated IQGAP1 适用: 人 WB, FACS 宿主: 兔 Monoclonal 25GB1900 unconjugated Recombinant Antibody
anti-IQ Motif Containing GTPase Activating Protein 1 (IQGAP1) (N-Term) antibodyIQGAP1 适用: 人 WB, ELISA 宿主: 兔 Polyclonal unconjugated
anti-IQ Motif Containing GTPase Activating Protein 1 (IQGAP1) antibodyIQGAP1 适用: 人 WB, IHC 宿主: 兔 Monoclonal unconjugated
anti-IQ Motif Containing GTPase Activating Protein 1 (IQGAP1) antibodyIQGAP1 适用: 人, 小鼠 WB, IHC, ELISA 宿主: 兔 Polyclonal unconjugated
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应用备注
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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. -
实验流程
- 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.
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限制
- 仅限研究用
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浓度
- 1.0 mg/mL
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缓冲液
- PBS, pH 7.2 containing 50 % Glycerol without preservatives.
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储存液
- Without preservative
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储存条件
- -20 °C
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储存方法
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Store the antibody (in aliquots) at -20 °C. Avoid repeated freezing and thawing.
Shelf life: one year from despatch. -
有效期
- 12 months
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- IQGAP1 (IQ Motif Containing GTPase Activating Protein 1 (IQGAP1))
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别名
- IQGAP1
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背景
- 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
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基因ID
- 8826
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UniProt
- P46940
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途径
- Signaling Events mediated by VEGFR1 and VEGFR2
抗原
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