ANTI-PHOSPHO LYN (TYR507) CLONE 1I11 ZOOMAB« RABBIT MONOCLONAL (C15-1669-330)

General description

ZooMAb^® antibodies represent an entirely new generation of recombinant monoclonal antibodies. Each ZooMAb^® antibody is manufactured using our proprietary recombinant expression system, purified to homogeneity, and precisely dispensed to produce robust and highly reproducible lot-to-lot consistency. Only top-performing clones are released for use by researchers. Each antibody is validated for high specificity and affinity across multiple applications, including its most commonly used application. ZooMAb^® antibodies are reliably available and ready to ship when you need them.

Specificity

Clone 1I11 is a ZooMAb^® rabbit recombinant monoclonal antibody that specifically detects Lyn protein kinase phosphorylated on tyrosine 507.

Immunogen

KLH-conjugated linear peptide corresponding to 10 amino acids surrounding phosphotyrosine 507 in human Lyn.

Application

Quality Control Testing

Evaluated by Western Blotting in Jurkat cell lysate.

Western Blotting Analysis): A 1:1,000 dilution of this antibody detected phospho Lyn (Tyr507) in Jurkat cell lysate.

Tested Applications

Western Blotting Analysis: A 1:1,000 dilution from a representative lot detected phospho Lyn (Tyr507) in Ramos cell lysate.

Peptide Inhibition Assay: Target band detection in a lysate from Jurkat and Ramos cells was prevented by preblocking of a representative lot with the immunogen phosphopeptide, but not the corresponding non-phosphopeptide.

Affinity Binding Assay: A representative lot of this antibody bound phospho Lyn (Tyr507) peptide with a KD of 5.8 x 10-6 in an affinity binding assay.

Immunocytochemistry Analysis: A 1:100 dilution from a representative lot detected phospho Lyn (Tyr507) in Jurkat cells.

Flow Cytometry Analysis: 1 µg from a representative lot detected phospho Lyn (Tyr507) in one million Jurkat cells.

Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user.

Target description

Tyrosine-protein kinase Lyn (UniProt: P07948; also known as EC:2.7.10.2, Lck/Yes-related novel protein tyrosine kinase, V-yes-1 Yamaguchi sarcoma viral related oncogene homolog, p53Lyn, p56Lyn) is encoded by the LYN (also known as JTK8) gene (Gene ID: 4067) in human. Lyn is a non-receptor protein tyrosine kinase of the Src family that plays a role in the regulation of innate and adaptive immune responses, hematopoiesis, responses to growth factors and cytokines, integrin signaling, and responses to DNA damage and genotoxic agents. It is widely expressed and is detected in epidermoid, hematopoietic, and neuronal cells. Its expression is also observed in primary neuroblastoma tumors. Two isoforms of Lyn (Lyn A and Lyn B) have been described that are produced by alternative splicing. They differ by a 20 amino acid region in their SH4 domain that encompasses pY32 motif. Activity of Lyn is regulated by protein interactions through its SH2/SH3 domains as well as via its phosphorylation. In its inactive state, it is phosphorylated at tyrosine 508 by C-terminal Src kinase that creates a binding site for its own SH2 domain. Its SH3 domain can bind an intramolecular Proline^®-motif situated between the SH2 domain (aa 129-226) and kinase domains (aa 247-501), helping generate a stabilized inactive kinase confirmation. Lyn is activated by dephosphorylation of the tyrosine 508 by CD45 and SHP-2 phosphatases. Activation can also occur via interactions with SH2 and/or SH3 domain binding motifs, which compete with Lyn′s own SH3/SH2 intramolecular interaction sites, thereby releasing the auto-inhibitory configuration of the kinase domain. Lyn can then trans-autophosphorylate at tyrosine 397 within the activation loop to generate a highly active enzyme. Lyn plays an important role in the regulation of B-cell differentiation, proliferation, survival, and apoptosis, and is also important for immune self-tolerance. It acts downstream of several immune receptors. Lyn is also reported to down-regulate signaling pathways by phosphorylation of immunoreceptor tyrosine-based inhibitory motifs (ITIM), that then serve as binding sites for phosphatases, such as SHP-1, and SHP-2, that modulate signaling by dephosphorylation of kinases and their substrates. This ZooMAb^® recombinant monoclonal antibody, generated by our propriety technology, offers significantly enhanced specificity, Affinity^™, reproducibility, and stability over conventional monoclonals.(Ref.: Ingley, E. (2012). Cell Commun. Signal. 10; 21; Xu, W., et al. (1999). Mol. Cell. 3(5); 629-638; Hirao, A., et al. (1997). EMBO J. 16(9); 1342-2351; Hata, A., et al. (1994). Mol. Cell Biol. 14(11); 7306-7313).

Physical form

Purified recombinant rabbit monoclonal antibody IgG, lyophilized in PBS, 5% Trehalose, normal appearance a coarse or translucent resin. The PBS/trehalose components in the ZooMAb formulation can have the appearance of a semi-solid (bead like gel) after lyophilization. This is a normal phenomenon. Please follow the recommended reconstitution procedure in the data sheet to dissolve the semi-solid, bead-like, gel-appearing material. The resulting antibody solution is completely stable and functional as proven by full functional testing. Contains no biocide or preservatives, such as azide, or any animal by-products. Larger pack sizes provided as multiples of 25 µL.

Reconstitution

300 µg/mL after reconstitution at 25 µL per vial. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.

Storage and Stability

Recommend storage of lyophilized product at 2-8°C; Before reconstitution, micro-centrifuge vials briefly to spin down material to bottom of the vial; Reconstitute each vial by adding 25 µL of filtered lab grade water or PBS; Reconstituted antibodies can be stored at 2-8°C, or -20°C for long term storage. Avoid repeated freeze-thaws.

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Legal Information

Affinity is a trademark of Mine Safety Appliances Co.

Proline is a registered trademark of Sartorius Biohit Liquid Handling Oy

ZooMAb is a registered trademark of Merck KGaA, Darmstadt, Germany

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.