Application
Lysates prepared from COS cells overexpressing human wild-type PAG3 alone (1) cotransfected with human wild-type PAG3 and mouse Pyk2 (2-5), or mutant [Y724F/Y763F] PAG3 and mouse Pyk2 (6) were resolved by SDS-PAGE on an 8% polyacrylamide gel and transferred to PVDF. Membranes were blocked with a 5% BSA-TBST buffer for one hour at room temperature, then incubated with PAG3 [pY724] antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1, 2, 6), the non-phosphopeptide corresponding to the immunogen (3), a generic phosphotyrosine-containing peptide (4), or, the phosphopeptide immunogen (5). After washing, membranes were incubated with goat F(ab)2 anti-rabbit IgG HRP conjugate in 3% BSA-TBST buffer, and bands were detected. The data show that only the peptide corresponding to PAG3 [pY724] blocks the signal, and the signal is absent in the mutant extracts, demonstrating the specificity of the antibody.| Product Name | PAG3 (phospho Tyr724) antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | Rabbit polyclonal to PAG3 (phospho Y724)PAG3 (paxillin-associated protein with ADP-ribosylation factor [ARF] GTPase-activating protein [GAP] activity, number 3) is a member of the ARF GAP family of proteins that associates with the cytoskeleton, plays an important role in mediating actin cytoskeletal organization in response to adhesion and growth factor stimulation, and is involved in paxillin subcellular localization. During monocyte maturation, the elevation of paxillin expression and integrin activation is associated with the induction of PAG3 expression. Overexpression of PAG3 inhibits paxillin recruitment to focal contacts and hinders cellular migration. PAG3 is phosphorylated by Pyk2 kinase on tyrosines 724 and 763. |
| Immunogen | Synthetic phosphopeptide derived from the region of human PAG3 that contains tyrosine 724. |
| Modification | p-Tyr724 |
| Clonality | Polyclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | WB |
| WB: 1/1000. Detects a band of approximately 160 kDa (predicted molecular weight: 112 kDa). Optimal dilutions/concentrations should be determined by the end user.: |
|
| Species Reactivity | HumanMouse |
| Concentration | 1mg/ml |
| Purification | Affinity purified |
| Gene Synonyms | Paxillin associated protein with ARFGAP activity 3 Pyk2 C terminus associated protein PAG3 Development and differentiation enhancing factor 2 PAP Paxillin associated protein with ARFGAP activity3 DDEF2 Development and differentiation enhancing fact |
|---|---|
| Target | PAG3 |
| Tissue Specificity | Lysates prepared from COS cells overexpressing human wild-type PAG3 and cotransfected with mouse Pyk2 were immunoblotted in the presence of non-phosphopeptide corresponding to the immunogen, a generic phosphotyrosine-containing peptide or the phosphopeptide immunogen. The data show that only the peptide corresponding to phospho Y724 PAG3 blocks the signal, and the signal is absent when mutant PAG3 extracts are used, demonstrating the specificity of the antibody. |
| Cellular Localization | Cytoplasmic and peripheral membrane protein associated with the plasma membrane and with Golgi stacks. Colocalizes with F actin and ARF6 in phagocytic cups. |
Application
Lysates prepared from COS cells overexpressing human wild-type PAG3 alone (1) cotransfected with human wild-type PAG3 and mouse Pyk2 (2-5), or mutant [Y724F/Y763F] PAG3 and mouse Pyk2 (6) were resolved by SDS-PAGE on an 8% polyacrylamide gel and transferred to PVDF. Membranes were blocked with a 5% BSA-TBST buffer for one hour at room temperature, then incubated with PAG3 [pY724] antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1, 2, 6), the non-phosphopeptide corresponding to the immunogen (3), a generic phosphotyrosine-containing peptide (4), or, the phosphopeptide immunogen (5). After washing, membranes were incubated with goat F(ab)2 anti-rabbit IgG HRP conjugate in 3% BSA-TBST buffer, and bands were detected. The data show that only the peptide corresponding to PAG3 [pY724] blocks the signal, and the signal is absent in the mutant extracts, demonstrating the specificity of the antibody.| Positive Control | COS cells overexpressing human wild type PAG3, cotransfected with mouse Pyk2 |
|---|---|
| Application Notes | WB: 1/1000. Detects a band of approximately 160 kDa (predicted molecular weight: 112 kDa). Optimal dilutions/concentrations should be determined by the end user.: |
| Form | Liquid |
|---|---|
| Storage Instructions | Keep as concentrated solution. Store at 4ºC short term. For extended storage aliquot and store at -20ºC or below. Avoid freeze-thaw cycles. |
| Storage Buffer | Preservative: 0.05% Sodium Azide. Constituents: 50% Glycerol, PBS (without Mg 2+ andCa 2+ ), 1mg/ml BSA. pH 7.3. |
