Anti-PAK1 + PAK2 + PAK3 (phospho S141) antibody (ab5247)
Key features and details
- Rabbit polyclonal to PAK1 + PAK2 + PAK3 (phospho S141)
- Suitable for: WB
- Reacts with: Recombinant fragment
- Isotype: IgG
Overview
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Product name
Anti-PAK1 + PAK2 + PAK3 (phospho S141) antibody
See all PAK1 + PAK2 + PAK3 primary antibodies -
Description
Rabbit polyclonal to PAK1 + PAK2 + PAK3 (phospho S141) -
Host species
Rabbit -
Tested Applications & Species
See all applications and species dataApplication Species WB Recombinant fragment
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Immunogen
Chemically synthesized phosphopeptide derived from a region of human PAK2 that contains serine 141. The sequence is conserved in rat and rabbit.
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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.
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General 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.
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, 0.1% BSA
BSA is IgG and protease free -
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
Images
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Western blot - Anti-PAK1 + PAK2 + PAK3 (phospho S141) antibody (ab5247)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
µ g/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.
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)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 0.50µ g/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.
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.
