Recombinant Anti-RhoGAP antibody [EP489Y] (ab32328)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP489Y] to RhoGAP
- Suitable for: WB, IHC-P, Flow Cyt (Intra)
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-RhoGAP antibody [EP489Y]
See all RhoGAP primary antibodies -
Description
Rabbit monoclonal [EP489Y] to RhoGAP -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, Flow Cyt (Intra)more details
Unsuitable for: ICC/IF or IP -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide within Human RhoGAP aa 1150-1250. The exact sequence is proprietary.
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Positive control
- Human stomach carcinoma and the following cell lysates: Hela, NIH 3T3, Jurkat, 293, K562, MCF-7, A431 and PC12.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP489Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab32328 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB | (1) |
1/500. Detects a band of approximately 190 kDa (predicted molecular weight: 171 kDa).
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IHC-P | (2) |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Flow Cyt (Intra) |
1/60 - 1/1000.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Notes |
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WB
1/500. Detects a band of approximately 190 kDa (predicted molecular weight: 171 kDa). |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Flow Cyt (Intra)
1/60 - 1/1000. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Target
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Relevance
The p190 RhoGAP protein is associated with p120 RasGAP in growth-factor stimulated and tyrosine kinase transformed cells. It functions as a GTPase - activating protein (GAP) for Rho and Rac family proteins, which are involved in regulating cytoskeletal actin and membrane ruffling. The antibody appears to couple signal transduction via Ras and Rho through its association with RasGAP. The presence of two adjacent SH2 domains in the p21 RAS GAP indicates that GAP might interact directly with tyrosine kinases. p190B can be directly phosphorylated on a single identified tyrosine residue by the activated insulin and IGF-1 receptors. This phosphorylation causes p190B to translocate to the lipid rafts of the plasma membrane, where it can facilitate the inactivation of the Rho GTPase. P190B also has been shown to be tyrosine phosphorylated by cSrc and vSrc. Additionally the GAP domain of p190B is shown to attenuate signal transducing activity of Rac, Rho and CDC42. -
Cellular localization
Cytoplasmic -
Database links
- Entrez Gene: 394 Human
- Entrez Gene: 11855 Mouse
- Entrez Gene: 299012 Rat
- Omim: 602680 Human
- SwissProt: Q13017 Human
- SwissProt: P97393 Mouse
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Alternative names
- ARHGAP 5 antibody
- ARHGAP5 antibody
- p190 B antibody
see all
Images
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Overlay histogram showing A431 cells stained with ab32328 (red line). The cells were fixed with 80% methanol (5 min)/ and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32328, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
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Anti-RhoGAP antibody [EP489Y] (ab32328) at 1/500 dilution + Mouse MEFs whole cell lysate at 50 µg
Secondary
HRP-conjugated Goat anti-rabbit IgG polyclonal at 1/2000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 171 kDa
Observed band size: 190 kDa why is the actual band size different from the predicted?
Exposure time: 5 minutes
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All lanes : Anti-RhoGAP antibody [EP489Y] (ab32328) at 1/500 dilution
Lane 1 : HeLa cell lysate
Lane 2 : Jurkat cell lysate
Lane 3 : MCF-7 cell lysate
Lane 4 : K562 cell lysate
Lane 5 : A431 cell lysate
Lane 6 : 293 cell lysate
Lane 7 : NIH 3T3 cell lysate
Lane 8 : PC12 cell lysate
Predicted band size: 171 kDa
Observed band size: 190 kDa why is the actual band size different from the predicted? -
ab32328, at a 1/100 dilution, staining human RhoGAP in stomach carcinoma by Immunohistochemistry, Paraffin embedded tissue
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
References (4)
ab32328 has been referenced in 4 publications.
- Yang C et al. Transcriptomic Analysis Identified ARHGAP Family as a Novel Biomarker Associated With Tumor-Promoting Immune Infiltration and Nanomechanical Characteristics in Bladder Cancer. Front Cell Dev Biol 9:657219 (2021). PubMed: 34307347
- Kim SR et al. Clinical Significance of CLDN18.2 Expression in Metastatic Diffuse-Type Gastric Cancer. J Gastric Cancer 20:408-420 (2020). PubMed: 33425442
- Zhu L et al. Impaired autophagic degradation of lncRNA ARHGAP5-AS1 promotes chemoresistance in gastric cancer. Cell Death Dis 10:383 (2019). PubMed: 31097692
- Repetto D et al. p140Cap regulates memory and synaptic plasticity through Src-mediated and citron-N-mediated actin reorganization. J Neurosci 34:1542-53 (2014). Mouse . PubMed: 24453341