Recombinant Anti-RAP1GAP antibody [Y134] (ab32373)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [Y134] to RAP1GAP
- Suitable for: Flow Cyt (Intra), ICC/IF, WB, IHC-P, IP
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-RAP1GAP antibody [Y134]
See all RAP1GAP primary antibodies -
Description
Rabbit monoclonal [Y134] to RAP1GAP -
Host species
Rabbit -
Specificity
ab32373 recognises Rap1 GTPase-activating protein 1 -
Tested applications
Suitable for: Flow Cyt (Intra), ICC/IF, WB, IHC-P, IPmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Jurkat, SH-SY5Y cell lysate, mouse and rat brain lysate IHC: Human breast cancer ICC/IF: HeLa cells IP: C6 cell lysate
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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. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
Y134 -
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 ab32373 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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Flow Cyt (Intra) |
1/30 - 1/50.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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ICC/IF |
1/100 - 1/250.
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WB |
1/10000 - 1/50000. Detects a band of approximately 95 kDa (predicted molecular weight: 73 kDa).
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IHC-P | (1) |
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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IP |
1/20 - 1/60.
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Notes |
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Flow Cyt (Intra)
1/30 - 1/50. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ICC/IF
1/100 - 1/250. |
WB
1/10000 - 1/50000. Detects a band of approximately 95 kDa (predicted molecular weight: 73 kDa). |
IHC-P
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
IP
1/20 - 1/60. |
Target
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Function
GTPase activator for the nuclear Ras-related regulatory protein RAP-1A (KREV-1), converting it to the putatively inactive GDP-bound state. -
Tissue specificity
Significant expression seen in the brain, kidney and pancreas. Abundant in the cerebral cortex and expressed at much lower levels in the spinal cord. Not detected in the lymphoid tissues. -
Sequence similarities
Contains 1 GoLoco domain.
Contains 1 Rap-GAP domain. -
Cellular localization
Golgi apparatus membrane. - Information by UniProt
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Database links
- Entrez Gene: 5909 Human
- Entrez Gene: 110351 Mouse
- Entrez Gene: 313644 Rat
- Omim: 600278 Human
- SwissProt: P47736 Human
- SwissProt: A2ALS5 Mouse
- Unigene: 148178 Human
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Alternative names
- KIAA0474 antibody
- Rap1 GTPase activating protein 1 antibody
- RAP1 GTPase activating protein antibody
see all
Images
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All lanes : Anti-RAP1GAP antibody [Y134] (ab32373) at 1/10000 dilution (purified)
Lane 1 : Mouse brain lysate
Lane 2 : Rat brain lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 73 kDa
Observed band size: 95 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBSTThe molecular weight observed is consistent with what has been described in the literature (PMID: 9346962, PMID: 15632203, PMID: 30144784).
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Anti-RAP1GAP antibody [Y134] (ab32373) at 1/10000 dilution (purified) + Jurkat cell lysate at 20 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 73 kDa
Observed band size: 95 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBSTThe molecular weight observed is consistent with what has been described in the literature (PMID: 9346962, PMID: 15632203, PMID: 30144784).
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Anti-RAP1GAP antibody [Y134] (ab32373) at 1/10000 dilution (purified) + SH-SY5Y cell lysate at 20 µg
Secondary
Anti-rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 73 kDa
Observed band size: 95 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBSTThe molecular weight observed is consistent with what has been described in the literature (PMID: 9346962, PMID: 15632203, PMID: 30144784).
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Anti-RAP1GAP antibody [Y134] (ab32373) at 1/50000 dilution (unpurified) + Jurkat cell lysate
Predicted band size: 73 kDa
Observed band size: 95 kDa why is the actual band size different from the predicted?The molecular weight observed is consistent with what has been described in the literature (PMID: 9346962, PMID: 15632203, PMID: 30144784).
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Immunohistochemical staining of paraffin embedded human cerebral cortex with purified ab32373 at a working dilution of 1/1000. The secondary antibody used is ab97051, a goat anti-rabbit IgG (H&L) at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
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Immunohistochemical staining of paraffin-embedded human breast cancer tissue using unpurified ab32373 at 1/100 dilution.
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Immunofluorescence staining of HeLa cells with purified ab32373 at a working dilution of 1/250, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4 % PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab32373 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.
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ICC/IF image of unpurified ab32373 stained HeLa 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 (ab32373, 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.
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Overlay histogram showing Jurkat cells fixed in 4% PFA and stained with purified ab32373 at a dilution of 1/30 (red line). The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit at a dilution of 1/500. Rabbit monoclonal IgG was used as an isotype control (black line) and cells incubated in the absence of both primary and secondary antibody were used as a negative control (blue line).
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Overlay histogram showing SH-SY5Y cells stained with unpurified ab32373 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab32373, 1/100) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.
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ab32373 (purified) at 1/20 immunoprecipitating RAP1GAP in 10 μg C6 cell lysate (Lanes 1 and 2, observed at 82-95 kDa). Lane 3 - Rabbit monoclonal IgG (ab172730). For western blotting, HRP Veriblot for IP Detection Reagent (ab131366) was used for detection (1/1000). Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST
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 (11)
ab32373 has been referenced in 11 publications.
- Gao Y et al. Rap1GAP Mediates Angiotensin II-Induced Cardiomyocyte Hypertrophy by Inhibiting Autophagy and Increasing Oxidative Stress. Oxid Med Cell Longev 2021:7848027 (2021). PubMed: 33936386
- Li H et al. Mex3a promotes oncogenesis through the RAP1/MAPK signaling pathway in colorectal cancer and is inhibited by hsa-miR-6887-3p. Cancer Commun (Lond) 41:472-491 (2021). PubMed: 33638620
- Mao X et al. ELK4-mediated lncRNA SNHG22 promotes gastric cancer progression through interacting with EZH2 and regulating miR-200c-3p/Notch1 axis. Cell Death Dis 12:957 (2021). PubMed: 34663788
- Zhang B et al. Arecoline Enhances Phosphodiesterase 4A Activity to Promote Transforming Growth Factor-β-Induced Buccal Mucosal Fibroblast Activation via cAMP-Epac1 Signaling Pathway. Front Pharmacol 12:722040 (2021). PubMed: 34819854
- Yan Z et al. Downregulation of Rap1GAP Expression Activates the TGF-β/Smad3 Pathway to Inhibit the Expression of Sodium/Iodine Transporter in Papillary Thyroid Carcinoma Cells. Biomed Res Int 2021:6840642 (2021). PubMed: 34840979