Phospho-Rac1+Cdc42 (S71) Recombinant Rabbit Monoclonal Antibody [JJ086-9]
Catalog# ET1701-40
Phospho-Rac1+Cdc42 (S71) Recombinant Rabbit Monoclonal Antibody [JJ086-9]
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WB
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IF-Cell
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IHC-P
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Human
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unconjugated
Overview
Product Name
Phospho-Rac1+Cdc42 (S71) Recombinant Rabbit Monoclonal Antibody [JJ086-9]
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic phospho-peptide corresponding to residues surrounding Ser71 of Human CDC42 aa 41-90 / 191.
Species Reactivity
Human
Validated Applications
WB, IF-Cell, IHC-P
Molecular Weight
Predicted band size: 21 kDa
Positive Control
A431, Hela, human lung carcinoma tissue, human liver carcinoma tissue.
Conjugation
unconjugated
Clone Number
JJ086-9
RRID
Product Features
Form
Liquid
Concentration
Storage Instructions
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
Storage Buffer
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
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WB
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1:500-1:1,000
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IF-Cell
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1:50-1:200
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IHC-P
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1:50-1:200
Target
Function
A large number of low molecular weight GTP-binding proteins of the Ras superfamily have been identified in eukaryotic cells; they regulate many fundamental processes such as cell growth, vesicle traffic and cytoskeletal organization. GTPase-activating proteins accelerate the intrinsic rate of GTP hydrolysis of Ras-related proteins, resulting in downregulation of their active form. Rac 1 is activated in a type I interferon (IFN) dependent manner; its function is required for downstream engagement of the p38 MAP kinase pathway. The p38 MAP kinase plays an essential role in IFN-dependent transcriptional regulation. The serine/threonine kinase Akt, of the phospho-inositide 3-kinase signal transduction pathway phosphorylates serine 71 of Rac 1. The superfamily of GTP-binding proteins, for which the Ras proteins are prototypes, has been implicated in regulation of diverse biological activities involving various aspects of cell growth and division.One mammalian member of the family, Cdc42, has an amino acid sequence that is similar to those of various members of the Ras superfamily proteins, including N-, K- and H-Ras, Rho proteins and the Rac proteins. On the basis of in vitro phosphorylation studies, it has been suggested that human Cdc42 may function in the signaling pathway of the EGF receptor or related growth factor receptor protein kinases. The Dbl oncogene has been shown to specifically catalyze dissociation of GDP from human Cdc42.
Background References
1. Gerasimcik N et al. The Rho GTPase Cdc42 Is Essential for the Activation and Function of Mature B Cells. J Immunol 194:4750-8 (2015).
2. Yoshida S et al. Growth factor signaling to mTORC1 by amino acid-laden macropinosomes. J Cell Biol 211:159-72 (2015).
Sequence Similarity
Belongs to the small GTPase superfamily. Rho family. CDC42 subfamily.
Post-translational Modification
(Microbial infection) AMPylation at Tyr-32 and Thr-35 are mediated by bacterial enzymes in case of infection by H.somnus and V.parahaemolyticus, respectively. AMPylation occurs in the effector region and leads to inactivation of the GTPase activity by preventing the interaction with downstream effectors, thereby inhibiting actin assembly in infected cells. It is unclear whether some human enzyme mediates AMPylation; FICD has such ability in vitro but additional experiments remain to be done to confirm results in vivo.; Phosphorylated by SRC in an EGF-dependent manner, this stimulates the binding of the Rho-GDP dissociation inhibitor RhoGDI.; (Microbial infection) Glycosylated at Tyr-32 by Photorhabdus asymbiotica toxin PAU_02230. Mono-O-GlcNAcylation by PAU_02230 inhibits downstream signaling by an impaired interaction with diverse regulator and effector proteins of CDC42 and leads to actin disassembly.
Subcellular Location
Cell junction, Cell membrane, Cell projection, Cytoplasm, Membrane, Nucleus, Synapse.
Synonyms
CDC42Hs antibody
Cell division cycle 42 antibody
G25K antibody
MIG5 antibody
Migration inducing gene 5 antibody
p21 Rac1 antibody
Ras like protein TC25 antibody
Small GTP binding protein Cdc42 antibody
TC25 antibody
Images
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☑ Cell treatment (CT)
Western blot analysis of Phospho-Rac1+Cdc42 (S71) on different lysates with Rabbit anti-Phospho-Rac1+Cdc42 (S71) antibody (ET1701-40) at 1/1,000 dilution.
Lane 1: A431 cell lysate
Lane 2: A431 treated with 100ng/mL EGF for 30 minutes cell lysate
Lane 3: MCF7 cell lysate
Lysates/proteins at 20 µg/Lane.
Predicted band size: 21 kDa
Observed band size: 25 kDa
Exposure time: 3 minutes; ECL: K1802;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1701-40) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
ICC staining of Phospho-Rac1+Cdc42 (S71) in A431 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1701-40, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
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ICC staining of Phospho-Rac1+Cdc42 (S71) in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1701-40, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
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Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue using anti-Phospho-Rac1+Cdc42 (S71) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-40, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
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Immunohistochemical analysis of paraffin-embedded human liver carcinoma tissue using anti-Phospho-Rac1+Cdc42 (S71) antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-40, 1/50) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"