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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| H, R, M, B, Av | ICC, IP, WB | M | Purified | Monoclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | 05-417 |
| Replaces | 04-581 |
| Brand Family | Upstate |
| Trade Name |
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| Description | Anti-Paxillin Antibody, clone 5H11 |
| Alternate Names |
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| Background Information | Paxillin, a focal adhesion scaffolding protein, is phosphorylated on multiple tyrosine residues in response various stimulations including integrin stimulation, growth factor stimulation, and oncogenic transformation (v-Src, v-Crk and BCR/ABL). Paxillin binds many other focal adhesion proteins including FAK, Vinculin, Talin, and Tensin. As such, Paxillin and it tyrosine phosphorylation, help regulate the dynamics of the focal adhesions and the actin cytoskeleton which results in cell morphology and cell migration characteristics of the cell. Paxillin is most notable phosphorylated on tyrosines 31 and 118 (Tyr31 and Tyr118). Interestingly, Vinculin and FAK bind to overlapping sites on Paxillin. Additionally, there is an increase in Paxillin phosphorylation at Tyr118 in vinculin (-/-) and Vinculin Y822F cells. Paxillin Y31F Y118F expression restores apoptosis and inhibits Erk activity. |
| Product Information | |
|---|---|
| Format | Purified |
| Control |
|
| Presentation | Purified mouse monoclonal IgG1 in buffer containing 0.1 M Tris-glycine, pH 7.4, 0.15 M NaCl, 0.05% sodium azide. Store at -20°C. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | Anti-Paxillin Antibody, clone 5H11 is an antibody against Paxillin for use in IC, IP & WB. |
| Key Applications |
|
| Application Notes | Immunoprecipitation: 4 μg of a previous lot of antibody immunoprecipitated paxillin from 500 μg of a 3T3/A31 RIPA lysate. Immunocytochemistry: 5-10 μg/mL of a previous lot gave positive immunostaining for paxillin in REF (rat embryo fibroblast) 52 cells fixed with 3.7% paraformaldehyde for 5 minutes. |
| Biological Information | |
|---|---|
| Immunogen | GST fusion protein containing fulllength human paxillin. Clone 5H11. |
| Clone | 5H11 |
| Concentration | Please refer to the Certificate of Analysis for the lot-specific concentration. |
| Host | Mouse |
| Specificity | This antibody recognizes paxillin. |
| Isotype | IgG1 |
| Species Reactivity |
|
| Species Reactivity Note | Human, mouse, rat, chicken and bovine. Other species cross-reactivity not tested. |
| Antibody Type | Monoclonal Antibody |
| Entrez Gene Number |
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| Gene Symbol |
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| Purification Method | Protein G Purified |
| UniProt Number |
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| UniProt Summary | FUNCTION: SwissProt: P49023 # Cytoskeletal protein involved in actin-membrane attachment at sites of cell adhesion to the extracellular matrix (focal adhesion). SIZE: 591 amino acids; 64533 Da SUBUNIT: Binds in vitro to vinculin as well as to the SH3 domain of c-SRC and, when tyrosine phosphorylated, to the SH2 domain of V-CRK. Isoform beta binds to focal adhesion kinase but weakly to vinculin. Isoform gamma binds to vinculin but only weakly to focal adhesion kinase. Interacts with GIT1, NUDT16L1/SDOS, PARVA and TGFB1I1. Component of cytoplasmic complexes, which also contain GIT1, ARHGEF6 and PAK1 (By similarity). Binds DDEF2. Interacts with unphosphorylated ITGA4. Interacts with RNF5. PTM: Phosphorylated on tyrosine residues during integrin-mediated cell adhesion, embryonic development, fibroblast transformation and following stimulation of cells by mitogens. SIMILARITY: SwissProt: P49023 ## Belongs to the paxillin family. & Contains 4 LIM zinc-binding domains. |
| Molecular Weight | 68 kDa |
| Product Usage Statements | |
|---|---|
| Quality Assurance | Routinely evaluated by Western Blot on RIPA lysates from 3T3/A31 fibroblasts. Western Blot Analysis: 0.5-2 μg/mL of this lot detected paxillin in RIPA lysates from 3T3/A31 fibroblasts. |
| Usage Statement |
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| Storage and Shipping Information | |
|---|---|
| Storage Conditions | Stable for 1 year at -20ºC from date of receipt. Handling Recommendations: Upon first thaw, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. Note: Variability in freezer temperatures below -20°C may cause glycerol-containing solutions to become frozen during storage. |
| Packaging Information | |
|---|---|
| Material Size | 250 µg |