Phospho-Syk (Tyr348) Monoclonal Antibody (moch1ct), PE

货号：12-9014-41,12-9014-42

规格：25 tests,100 tests

价格：1340,3202

产品类型：流式抗体

品牌：eBioscience

物种：人/小鼠

宿主：小鼠

抗体亚型：IgG

克隆号：moch1ct

荧光染料：PE

类型:	单抗	同型对照：	Mouse IgG1 kappa Isotype Control (P3.6.2.8.1), PE
浓度：	5 µL/Test	用法：	5 µL (0.06 µg)/test(Flow)

产品详细信息Description: This moch1ct monoclonal antibody recognizes human and mouse spleen tyrosine kinase (also known as SYK) when phosphorylated on tyrosine 348 (Y348). SYK is the founding member of the SYK family of kinases that also includes ZAP-70 (zeta-associated protein of 70 kD) and is expressed in hematopoietic cells, including B lymphocytes, immature (CD4, CD8 double-negative and double-positive) thymocytes, and myeloid cells, epithelial cell lines, and normal breast tissue. SYK is critical for Bcell receptor (BCR) signaling and B cell development. Autophosphorylation of at Y348 is necessary for SYK to become fully catalytically active and creates a docking site for the SH2 domains of Vav, Grb2, p85 subunit of PI3 kinase, and PLC gamma.Specificity of this moch1ct clone was confirmed by ELISA, flow cytometry, and western blotting.Applications Reported: This moch1ct antibody has been reported for use in intracellular staining followed by flow cytometric analysis.Applications Tested: This moch1ct antibody has been pre-titrated and tested by intracellular staining followed by flow cytometric analysis of stimulated mouse splenocytes. This can be used at 5 µL (0.06 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.Staining Protocol: All protocols work well for this monoclonal antibody. Use of Protocol A: Two-step protocol: intracellular (cytoplasmic) proteins allows for the greatest flexibility for detection of surface and intracellular (cytoplasmic) proteins. Use of Protocol B: One-step protocol: intracellular (nuclear) proteins is recommended for staining of transcription factors in conjunction with surface and phosphorylated intracellular (cytoplasmic) proteins. Protocol C: Two-step protocol: Fixation/Methanol allows for the greatest discrimination of phospho-specific signaling between unstimulated and stimulated samples, but with limitations on the ability to stain specific surface proteins (refer to "Clone Performance Following Fixation/Permeabilization" located in the Best Protocols Section under the Resources tab online). All Protocols can be found in the Flow Cytometry Protocols: "Staining Intracellular Antigens for Flow Cytometry Protocol" located in the Best Protocols Section under the Resources tab online.Excitation: 488-561 nm; Emission: 578 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser.Filtration: 0.2 µm post-manufacturing filtered.靶标信息Syk is a cytoplasmic protein tyrosine kinase that translocates to the plasma membrane upon B cell antigen receptor (BCR) or the high-affinity IgE receptor (FcepsilonRI) triggering, and phosphorylates downstream adaptor proteins, thereby providing docking sites for initiation of subsequent signaling pathways, such as calcium mobilization, cytoskeleton remodeling, or transcription of specific genes. Syk binds to the receptor assemblies through interactions of its pair of SH2 domains with ITAM motives of the receptor, which have been phosphorylated by Src-family kinases. These kinases also help to activate Syk by phosphorylation of its activation loop.

数据

Phospho-Syk (Tyr348) Antibody (12-9014-41) in FlowTOP: Intracellular staining of unstimulated (left) or 2-minute F (ab')2 Anti-Mouse IgM, u chain specific Functional Grade Purified-stimulated (Product # 16-5092-85) (right) mouse spleen cells with Anti-Mouse CD45R (B220) APC (Product # 17-0452-82) and Anti-Human/Mouse phospho-SYK (Y348) PE using the Intracellular Fixation and Permeabilization Buffer Set (Product # 88-8824-00) and protocol. BOTTOM: Intracellular staining of unstimulated (orange histogram), 2-minute F (ab')2 Anti-Mouse IgM, u chain specific Functional Grade Purified-stimulated (Product # 16-5092-85) (purple histogram), or hydrogen peroxide-activated sodium pervanadate-treated mouse spleen cells (green histogram) with Anti-Human/Mouse phospho-SYK (Y348) PE. CD45R (B220)+ lymphocytes were used for analysis.

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