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Products:Cell Biology >> Apoptosis >> Intracellular >> Kinases
Overview
Product name
Anti-PAK1 + PAK2 + PAK3 (phospho S141) antibody
See all PAK1 + PAK2 + PAK3 products (8) ...
Description
Rabbit polyclonal to PAK1 + PAK2 + PAK3 (phospho S141)
Tested applications
Cross reactivity
Reacts with
Mouse, Human
Predicted to work with
Rat, Rabbit
Immunogen
Chemically synthesized phosphopeptide derived from a region of human PAK2 that contains serine 141. The sequence is conserved in rat and rabbit.
Positive control
Cos cells transiently transfected with PAK 1, PAK 2, or PAK 3, and co transfected with V12 Cdc42 and grown either as adherent cultures to activate PAK, or placed in suspension to suppress PAK activation.
Properties
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
Preservative: 0.05% Sodium Azide
Constituents: PBS, 1mg/ml BSA (IgG, protease free). pH 7.3
Concentration
Concentration information loading...
Purity
Immunogen affinity purified
Purification notes
Purified from rabbit serum by sequential epitope-specific chromatography. 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 PAK. The final product is generated by affinity chromatography using a PAK 2 derived peptide that is phosphorylated at serine 141.
Primary antibody notes
P21-activated kinase (PAK) is actually a family of serine/threonine protein kinases, members of which are activated by small molecular weight GTPases. The three most common isoforms are PAK 1, PAK 2, and PAK 3 (also known as alpha PAK, gamma PAK, and beta PAK, respectively). These kinases contain numerous regulatory elements that trigger diverse signaling processes such as those initiated by activated GTPases, interaction with Src homology 3 (SH3) domains, and caspase mediated proteolytic cleavage. Autophosphorylation of serine 141 (serine 144 for PAK 1 and serine 139 PAK 3), catalyzed by Cdc42, is required for activation of PAK.
Clonality
Polyclonal
Isotype
IgG
Research areas
Cancer >> Signal transduction >> Protein phosphorylation >> Serine/threonine kinases >> Other
Signal Transduction >> Protein Phosphorylation >> Ser / Thr Kinases >> Other Kinases
Signal Transduction >> Cytoskeleton / ECM >> Extracellular Matrix >> Structures >> Focal Adhesions
Cell Biology >> Apoptosis >> Intracellular >> Kinases
Applications
Show applications key
Our Abpromise guarantee covers the use of ab5247 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
WB: Use a concentration of 0.1 - 1.0 µg/ml.Predicted molecular weight: 65 kDa.
Target
Relevance
In mammals, there are several identified isoforms of p21 Activated Protein Kinases or PAKs: PAK1 and PAK3 are mostly brain specific, while PAK2 is expressed ubiquitously. Mutations of the gene coding for PAK3 are associated with X linked mental retardation and PAK3 is a key regulator of synapse formation and plasticity in the hippocampus. PAK3 is thought to play a key role in regulation of cell shape and motility as well as cell death.
Cellular localization
Cytoplasm. Recruited to focal adhesions upon activation
Alternative names
- Alpha PAK antibody
- Beta PAK antibody
- Gamma PAK antibody
see all
Database links
- Entrez Gene: 5058 Human
- Entrez Gene: 5062 Human
- Entrez Gene: 5063 Human
- Entrez Gene: 18479 Mouse
- Entrez Gene: 18481 Mouse
- Entrez Gene: 224105 Mouse
- Entrez Gene: 29431 Rat
- Entrez Gene: 29432 Rat
see all
Anti-PAK1 + PAK2 + PAK3 (phospho S141) antibody images:
Western blot - Anti-PAK1 + PAK2 + PAK3 (phospho S141) antibody (ab5247)
Predicted band size : 65 kDa
Induction of Activation: Extracts prepared from Cos cells transiently transfected with PAK 1 and V12 Cdc42, and kept in suspension (inactive) or as adherent cultures (active) were resolved by SDSPAGE on a 10% Tris-glycine gel and transferred to PVDF. The membrane was either not treated or treated with lambda phosphatase, blocked with a 5% BSA-TBST buffer overnight at 4°C, and then incubated with 0.50 µg/mL ab5247 antibody for two hours at room temperature in a 3% BSA-TBST buffer. After washing, membranes were incubated with goat F(ab’)2 anti-rabbit IgG alkaline phosphatase and signals were detected using the Tropix WesternStar method.
Western blot - Anti-PAK1 + PAK2 + PAK3 (phospho S141) antibody (ab5247)
Predicted band size : 65 kDa
Peptide Competition: Extracts prepared from Cos cells transiently transfected with PAK 1 (lanes 1-4), PAK 2 (lanes 5-8), or PAK 3 (lanes 9-12) and cotransfected with V12 Cdc42 to induce activation were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF. Membranes were blocked with a 5% BSA-TBST buffer overnight at 4°C, then were incubated with 0.50 µg/mL ab5247 antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1, 5, 9), the nonphosphopeptide corresponding to the immunogen (2, 6, 10), a generic phosphoserine containing peptide (3, 7, 11), or, the phosphopeptide immunogen (4, 8, 12). After washing, membranes were incubated with goat F(ab’)2 anti-rabbit IgG alkaline phosphatase and signals were detected using the Tropix WesternStar method. The data show that the ab5247 antibody is highly selective for the phosphorylated form of PAK, and that the antibody can be used to detect an active form of PAK 1 as well as PAK 2 and PAK 3.
References for Anti-PAK1 + PAK2 + PAK3 (phospho S141) antibody (ab5247)
This product has been referenced in:
- Awasthi Aet al. Rap1b facilitates NK cell functions via IQGAP1-mediated signalosomes. J Exp Med 207:1923-38 (2010). WB; Mouse.Read more (PubMed: 20733035) »
See 1 publication for this product
Publishing research using ab5247? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"