Anti-RAC1 + Cdc42 (phospho S71) antibody (ab5482)
Key features and details
- Rabbit polyclonal to RAC1 + Cdc42 (phospho S71)
- Suitable for: IHC-P, WB
- Reacts with: Human
- Isotype: IgG
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Overview
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Product name
Anti-RAC1 + Cdc42 (phospho S71) antibody
See all RAC1 + Cdc42 primary antibodies -
Description
Rabbit polyclonal to RAC1 + Cdc42 (phospho S71) -
Host species
Rabbit -
Specificity
Cdc 42 [pS71] (100% homologous) and Rho A/B/C [pS73] (92% homologous) are expected to react. -
Tested applications
Suitable for: IHC-P, WBmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide corresponding to Human RAC1 + Cdc42 (phospho S71). The sequence is conserved in human and mouse RAC 1, 2, and 3, and Cdc 42 human, mouse, rat, dog and frog.
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Positive control
- WB: A431 cells treated with EGF. IHC-P: human skin, human stomach tissues
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General notes
RAC, Cdc 42 and Rho A, B, and C are members of a small RhoGTPase family that bind and hydrolyze GTP. GTP bound RAC 1 and cdc 42 play a pivotal role in controlling cell shape, adhesion, growth and transformation. Active Rac 1 is implicated in regulating serum response element (SRE), NFAT 1 and nuclear factor kappa B (NF kappa B) transcription activities. Activated RAC 1 and Cdc 42 bind and activate PAK 1, which in turn activates key downstream signaling proteins including MEKK 1 and JNK. RAC 1 and Cdc 42 are phosphorylated on serine 71, a putative Akt site located between the protein binding domain and GTP binding domain. Phosphorylation of RAC 1 on serine 71 regulates its GTP binding and GTPase activity.
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Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles. -
Storage buffer
pH: 7.30
Preservative: 0.05% Sodium azide
Constituents: PBS, 50% Glycerol (glycerin, glycerine), 0.1% BSA
BSA is IgG and protease free -
Concentration information loading...
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Purity
Immunogen affinity purified -
Purification notes
The antibody has been negatively preadsorbed using a non-phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non-phosphorylated RAC 1. The final product is generated by affinity chromatography using a RAC 1 derived peptide that is phosphorylated at serine 71. -
Primary antibody notes
RAC, Cdc 42 and Rho A, B, and C are members of a small RhoGTPase family that bind and hydrolyze GTP. GTP bound RAC 1 and cdc 42 play a pivotal role in controlling cell shape, adhesion, growth and transformation. Active Rac 1 is implicated in regulating serum response element (SRE), NFAT 1 and nuclear factor kappa B (NF kappa B) transcription activities. Activated RAC 1 and Cdc 42 bind and activate PAK 1, which in turn activates key downstream signaling proteins including MEKK 1 and JNK. RAC 1 and Cdc 42 are phosphorylated on serine 71, a putative Akt site located between the protein binding domain and GTP binding domain. Phosphorylation of RAC 1 on serine 71 regulates its GTP binding and GTPase activity. -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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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 ab5482 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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IHC-P |
1/20 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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WB | (1) |
1/1000. Detects a band of approximately 23 kDa.
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Notes |
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IHC-P
1/20 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
WB
1/1000. Detects a band of approximately 23 kDa. |
Target
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Relevance
Cdc42/Rac belongs to the superfamily of small GTPases that are structurally linked to the proto-oncogene product p21ras and are important for the control of cell growth and differentiation as well as for intracellular organization. Cdc42/Rac is an important upstream regulator of the protein kinase cascade that controls the SAPK/JNK and p38 activity. Recent data also suggest that constitutive active forms of Cdc42 can induce apoptosis through a mechanism requiring signaling through SAPK/JNK. -
Cellular localization
Cell membrane; Lipid anchor; Cytoplasmic side. -
Database links
- Entrez Gene: 5879 Human
- Entrez Gene: 998 Human
- Omim: 116952 Human
- Omim: 602048 Human
- SwissProt: P60953 Human
- SwissProt: P63000 Human
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Alternative names
- CDC42Hs antibody
- Cell division cycle 42 antibody
- G25K antibody
see all
Images
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Peptide Competition and Phosphatase Treatment:
Lysates prepared from A431 cells left unstimulated (1) or stimulated with EGF (2-6) were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes were either not treated (1-5) or treated with lambda phosphatase (6), blocked with a 5% BSA-TBST buffer for one hour at room temperature, and incubated with ab5482 antibody for two hours at room temperature in a 3% BSATBST buffer, following prior incubation with: no peptide (1, 2, 6), the nonphosphopeptide corresponding to the immunogen (3), a generic phosphoserine containing peptide (4), or, the phosphopeptide immunogen (5), After washing, membranes were incubated with goat F(ab’)2 anti-rabbit IgG HRP conjugate and bands were detected using the Pierce SuperSignal method. The data show that the peptide corresponding to ab5482 blocks the antibody signal. The data also shows that phosphatase stripping eliminates the signal, verifying that the antibody is pho -
Immunohistochemiscal analysis of paraffin-embedded human skin tissue labeling RAC1 + Cdc42 (phospho S71) with ab5482 at 1/100 dilution (right) compared to a negative control without primary antibody (left).
To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with ab5482 diluted in 3% BSA-PBS at a dilution of 1/100 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.
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Immunohistochemical analysis of paraffin-embedded human stomach tissue labeling RAC1 + Cdc42 (phospho S71) with ab5482 at 1/20 dilution (right) compared to a negative control without primary antibody (left).
To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with ab5482 diluted in 3% BSA-PBS at a dilution of 1/20 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (7)
ab5482 has been referenced in 7 publications.
- Li N et al. LPM580098, a Novel Triple Reuptake Inhibitor of Serotonin, Noradrenaline, and Dopamine, Attenuates Neuropathic Pain. Front Pharmacol 10:53 (2019). PubMed: 30837867
- Kang DS et al. Netrin-1/DCC-mediated PLC?1 activation is required for axon guidance and brain structure development. EMBO Rep 19:N/A (2018). PubMed: 30224412
- Giralt A et al. Pyk2 is essential for astrocytes mobility following brain lesion. Glia 64:620-34 (2016). PubMed: 26663135
- Maurya VK et al. Expression and activity of Rac1 is negatively affected in the dehydroepiandrosterone induced polycystic ovary of mouse. J Ovarian Res 7:32 (2014). PubMed: 24628852
- Haas S et al. DIGE proteome analysis reveals suitability of ischemic cardiac in vitro model for studying cellular response to acute ischemia and regeneration. PLoS One 7:e31669 (2012). WB ; Mouse . PubMed: 22384053
- Pickl M & Ries CH Comparison of 3D and 2D tumor models reveals enhanced HER2 activation in 3D associated with an increased response to trastuzumab. Oncogene 28:461-8 (2009). WB ; Human . PubMed: 18978815
- Tan AM et al. Neuropathic pain memory is maintained by Rac1-regulated dendritic spine remodeling after spinal cord injury. J Neurosci 28:13173-83 (2008). WB ; Rat . PubMed: 19052208