Phospho-LCK (Tyr505) Monoclonal Antibody (SRRCHA), eFluor 660

货号：50-9076-42

规格：100 tests

价格：3301

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：SRRCHA

荧光染料：eFluor 660

类型:	单抗	同型对照：	Mouse IgG1 kappa Isotype Control (P3.6.2.8.1), eFluor 660
浓度：	5 µL/Test	用法：	5 µL (0.125 µg)/test(Flow)

产品详细信息Description: This SRRCHA monoclonal antibody recognizes human and mouse LCK when phosphorylated on tyrosine 505 (Y505). Y505 is a negative-regulatory site that is phosphorylated by the kinase CSK. LCK is expressed in T cells where it associates with the cytoplasmic tails of the CD4 and CD8 co-receptors on T helper cells and cytotoxic T cells, respectively. When the TCR is engaged, LCK acts to phosphorylate the intracellular chains of CD3 and zeta chains of the TCR complex, allowing the cytoplasmic tyrosine kinase, ZAP-70, to bind to them. LCK then phosphorylates and activates ZAP-70, which then phosphorylates the adaptor protein, LAT, that serves as a docking site for a number of other proteins, including SLP-76, Grb2, and phospholipase C gamma1. Mice lacking expression of LCK show defects in T cell development and function.Applications Reported: This SRRCHA antibody has been reported for use in intracellular staining followed by flow cytometric analysis.Applications Tested: This SRRCHA antibody has been pre-titrated and tested by intracellular staining followed by flow cytometric analysis of stimulated normal human peripheral blood cells. This can be used at 5 µL (0.125 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.eFluor® 660 is a replacement for Alexa Fluor® 647. eFluor® 660 emits at 659 nm and is excited with the red laser (633 nm). Please make sure that your instrument is capable of detecting this fluorochome.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: 633-647 nm; Emission: 668 nm; Laser: Red Laser.Filtration: 0.2 µm post-manufacturing filtered.靶标信息LCK is a member of the Src-family tyrosine kinase, which are essential for signaling through the T-cell receptor (TCR) complex. Upon TCR triggering, LCK phosphorylates the ITAM motives in its zeta subunits establishing binding sites for the SH2 domains of the tyrosine kinase ZAP70, which is also phosphorylated by LCK and activated to generate subsequent signaling platforms by phosphorylation of adaptor LAT. The majority of LCK is localized to the plasma membrane, however, there is also a significant fraction associated with the Golgi apparatus which may contribute to Raf activation under conditions of weak stimulation through the TCR. LCK contains N-terminal sites for myristylation and palmitylation, a PTK domain, and SH2 and SH3 domains which are involved in mediating protein-protein interactions with phosphotyrosine-containing and proline-rich motifs, respectively. LCK is also involved in the regulation of apoptosis induced by various stimuli, but not by the death receptors. Diseases associated with LCK include Immunodeficiency 22 and CD45 deficiency. Alternatively splice variants of the LCK gene encoding different isoforms of the protein have been found.

数据

Phospho-LCK (Tyr505) Antibody (50-9076-41) in FlowTOP: Intracellular staining of untreated (left) or 5-minute hydrogen peroxide-activated sodium pervanadate-treated (right) normal human peripheral blood cells with Atni-Human CD3 PE (Product # 12-0037-42) and Anti-Human/Mouse phospho-LCK (Y505) eFluor® 660, using the Intracellular Fixation and Permeabilization Buffer Set (Product # 88-8824-00) and protocol. Cells in the lymphocytes gate were used for analysis. BOTTOM: Intracellular staining of untreated (orange histogram) or 5-minute hydrogen peroxide-activated sodium pervanadate-treated (purple histogram) normal human peripheral blood cells with Anti-Human CD3 PE (Product # 12-0037-42), Anti-Human CD19 FITC (Product # 11-0199-42), and Anti-Human/Mouse phospho-LCK (Y505) eFluor® 660, using the Intracellular Fixation and Permeabilization Buffer Set (Product # 88-8824-00) and protocol. Lymphocytes in the CD3+ (left) or CD19+ (right) gates were used for analysis.

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