Anti-IQGAP1 antibody (ab110203)
Key features and details
- Rabbit polyclonal to IQGAP1
- Suitable for: ICC/IF, WB, IHC-P
- Knockout validated
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-IQGAP1 antibody
See all IQGAP1 primary antibodies -
Description
Rabbit polyclonal to IQGAP1 -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species ICC/IF HumanIHC-P HumanWB Human
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Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Human placenta tissue lysate and HeLa and HEK293 whole cell lysates. IHC-P: Human skin melanoma tissue. ICC/IF: MCF7 cells.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab110203 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
GuaranteedTested applications are guaranteed to work and covered by our Abpromise guarantee.
PredictedPredicted to work for this combination of applications and species but not guaranteed.
IncompatibleDoes not work for this combination of applications and species.
Application Species ICC/IF HumanIHC-P HumanWB HumanAll applications DogChimpanzeeMacaque monkeyGorillaOrangutanApplication Abreviews Notes ICC/IF Use a concentration of 5 µg/ml.WB Use a concentration of 1 µg/ml. Detects a band of approximately 205 kDa (predicted molecular weight: 189 kDa).IHC-P Use a concentration of 1 µg/ml.Notes ICC/IF
Use a concentration of 5 µg/ml.WB
Use a concentration of 1 µg/ml. Detects a band of approximately 205 kDa (predicted molecular weight: 189 kDa).IHC-P
Use a concentration of 1 µg/ml.Target
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Function
Binds to activated CDC42 but does not stimulate its GTPase activity. It associates with calmodulin. Could serve as an assembly scaffold for the organization of a multimolecular complex that would interface incoming signals to the reorganization of the actin cytoskeleton at the plasma membrane. May promote neurite outgrowth. -
Tissue specificity
Expressed in the placenta, lung, and kidney. A lower level expression is seen in the heart, liver, skeletal muscle and pancreas. -
Sequence similarities
Contains 1 CH (calponin-homology) domain.
Contains 4 IQ domains.
Contains 1 Ras-GAP domain.
Contains 1 WW domain. -
Domain
Regions C1 and C2 can either interact with nucleotide-free CDC42, or interact together, depending on the phosphorylation state of Ser-1443. When Ser-1443 is not phosphorylated, C1 and C2 interact, which prevents binding of nucleotide-free CDC42 and promotes binding of GTP-bound CDC42. Phosphorylation of Ser-1443 prevents interaction between C1 and C2, which opens the structure of the C-terminus and allows binding and sequestration of nucleotide-free CDC42 on both C1 and C2. -
Post-translational
modificationsPhosphorylation of Ser-1443 by PKC prevents interaction between C1 and C2, allowing binding of nucleotide-free CDC42. Ser-1443 phosphorylation enhances the ability to promote neurite outgrowth. -
Cellular localization
Cell membrane. - Information by UniProt
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Database links
- Entrez Gene: 8826 Human
- Omim: 603379 Human
- SwissProt: P46940 Human
- Unigene: 430551 Human
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Alternative names
- HUMORFA01 antibody
- IQ motif containing GTPase activating protein 1 antibody
- IQGA1_HUMAN antibody
see all
Images
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Western blot - Anti-IQGAP1 antibody (ab110203)
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: IQGAP1 knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: HEK293 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab110203 observed at 190 kDa. Red - loading control, ab7291, observed at 52 kDa.
ab110203 was shown to specifically react with IQGAP1 when IQGAP1 knockout samples were used. Wild-type and IQGAP1 knockout samples were subjected to SDS-PAGE. ab110203 and ab7291 (loading control to alpha tubulin) were diluted 1 μg/mL and 1/2000 respectively and incubated overnight at 4°C. Blots were developed withGoat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging. -
Immunocytochemistry/ Immunofluorescence - Anti-IQGAP1 antibody (ab110203)ICC/IF image of ab110203 stained MCF7 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab110203 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% foarmaldehyde fixed (10 min) Hek293 cells at 1µg/ml, and in 100% methanol fixed (5 min) HeLa and MCF7 cells at 1µg/ml.
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Western blot - Anti-IQGAP1 antibody (ab110203)All lanes : Anti-IQGAP1 antibody (ab110203) at 1 µg/ml
Lane 1 : Human placenta tissue lysate - total protein (ab29745)
Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 3 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 189 kDa
Observed band size: 205 kDa why is the actual band size different from the predicted?
Additional bands at: 45 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 1 minute -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IQGAP1 antibody (ab110203)IHC image of IQGAP1 staining in Human skin melanoma formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab110203, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Protocols
Datasheets and documents
References (1)
ab110203 has been referenced in 1 publication.
- Müller MT et al. Interaction of microtubules and actin during the post-fusion phase of exocytosis. Sci Rep 9:11973 (2019). PubMed: 31427591
Images
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Western blot - Anti-IQGAP1 antibody (ab110203)
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: IQGAP1 knockout HAP1 cell lysate (20 µg)
Lane 3: HeLa cell lysate (20 µg)
Lane 4: HEK293 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab110203 observed at 190 kDa. Red - loading control, ab7291, observed at 52 kDa.
ab110203 was shown to specifically react with IQGAP1 when IQGAP1 knockout samples were used. Wild-type and IQGAP1 knockout samples were subjected to SDS-PAGE. ab110203 and ab7291 (loading control to alpha tubulin) were diluted 1 μg/mL and 1/2000 respectively and incubated overnight at 4°C. Blots were developed withGoat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging. -
Immunocytochemistry/ Immunofluorescence - Anti-IQGAP1 antibody (ab110203)
ICC/IF image of ab110203 stained MCF7 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab110203 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% foarmaldehyde fixed (10 min) Hek293 cells at 1µg/ml, and in 100% methanol fixed (5 min) HeLa and MCF7 cells at 1µg/ml.
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Western blot - Anti-IQGAP1 antibody (ab110203)All lanes : Anti-IQGAP1 antibody (ab110203) at 1 µg/ml
Lane 1 : Human placenta tissue lysate - total protein (ab29745)
Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 3 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 189 kDa
Observed band size: 205 kDa why is the actual band size different from the predicted?
Additional bands at: 45 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 1 minute
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IQGAP1 antibody (ab110203)
IHC image of IQGAP1 staining in Human skin melanoma formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab110203, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
