Invitrogen

Phospho-Syk (Tyr323, Tyr317) Polyclonal Antibody

This Antibody was verified by Cell treatment to ensure that the antibody binds to the antigen stated.

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Phospho-Syk (Tyr323, Tyr317) Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of Phospho-Syk pTyr323/pTyr317 was performed using 70% confluent log phase Jurkat cells treated with 100 uM H2O2 for 1 hour. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 1 hour at room temperature. The cells were labeled with Phospho-Syk pTyr323/pTyr317 Rabbit Polyclonal Antibody (Product # 44-234G) at 2 µg/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows untreated cells with no signal. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Phospho-Syk (Tyr323, Tyr317) Antibody in Western Blot (WB) — Upregulation, Antibody-Peptide Competition and Phosphatase Stripping. Extracts of Jurkat cells untreated (1) or treated with 10 mM H2O2 for 3 minutes (2-6) were resolved by SDS-PAGE on a 10% Tris-glycine gel and transferred to PVDF. The membrane was either left untreated (1-5) or treated with YOP phosphatase (6), then blocked with a 3% milk-TBST buffer for one hour at room temperature, and incubated with the Syk (pY323) antibody for two hours at room temperature in a 3% milk-TBST buffer, following prior incubation with: no peptide (1, 2, 6), the non-phosphopeptide corresponding to the phosphopeptide immunogen (3), a generic phosphotyrosine-containing peptide (4), or the phosphopeptide immunogen (5). After washing, the membrane was incubated with goat F (ab')2 anti-rabbit IgG HRP conjugate (Product # ALI4404) and signals were detected using the Pierce SuperSignal™ method. The data show that only the phosphopeptide corresponding to Syk (pY323) blocks the antibody signal, demonstrating the specificity of the antibody. The data also show the up-regulation of phosphorylation upon treatment with H2O2 in this cell system and that phosphatase stripping eliminates the signal, further verifying that the antibody is phospho-specific.

Phospho-Syk (Tyr323, Tyr317) Antibody — Modulation of expression of target protein by cell treatment demonstrates antibody specificity. Immunofluorescence analysis of Phospho-Syk (Tyr323, Tyr317) using Phospho-Syk (Tyr323, Tyr317) Rabbit Polyclonal Antibody (Product # 44-234G) shows cytoplasmic localization of Phospho-Aurora A (Thr288) in Jurkat cells treated with hydrogen peroxide. {TM}

产品信息

44-234G

应用	建议稀释比	已发表文章
免疫印迹 (WB)	1:1,000	查看1篇已发表文章 1篇已发表文章
免疫细胞化学 (ICC/IF)	1:250	-

产品规格
种属反应	Human, Mouse
已发表种属	Mouse
宿主/亚型	Rabbit / IgG
分类	Polyclonal
类型	Antibody
抗原	The antiserum was produced against a chemically synthesized phosphopeptide derived from a region of human Syk that contains tyrosine 323 (tyrosine 317 in the mouse sequence).
偶联物	Unconjugated
形式	Liquid
纯化类型	Antigen affinity chromatography
保存液	Dulbecco's PBS, pH 7.3, with 1mg/mL BSA
内含物	0.05% sodium azide
保存条件	-20°C
RRID	AB_2533612

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.

仅用于科研。不用于诊断过程。未经明确授权不得转售。

Product References

Immune Sensing of Cell Death through Recognition of Histone Sequences by C-Type Lectin-Receptor-2d Causes Inflammation and Tissue Injury.

Immunity

Lai JJ,Cruz FM,Rock KL

44-234G was used in Western Blot to examine the effects of histones binding to C-type lectin receptors following necrotic cell death.

Tue Jan 14 00:00:00 EST 2020

生物信息学

蛋白别名: DKFZp313N1010; EC 2.7.10.2; FLJ25043; FLJ37489; HGNC:11205; kinase Syk; KSYK; p72-Syk; SP1; Spleen tyrosine kinase; syk signaling; tyrosine-kinase syk; Tyrosine-protein kinase SYK

基因别名: p72-Syk; ptk72; SYK; Sykb

UniProt ID: (Human) P43405, (Mouse) P48025

Entrez Gene ID: (Human) 6850, (Mouse) 20963

分子生物学功能: kinase modulator

angiogenesis lymph vessel development positive regulation of receptor internalization stimulatory C-type lectin receptor signaling pathway adaptive immune response macrophage activation involved in immune response neutrophil activation involved in immune response leukocyte activation involved in immune response serotonin secretion by platelet protein phosphorylation inflammatory response leukocyte cell-cell adhesion transmembrane receptor protein tyrosine kinase signaling pathway integrin-mediated signaling pathway activation of JUN kinase activity cell proliferation organ morphogenesis regulation of platelet activation regulation of tumor necrosis factor-mediated signaling pathway viral process peptidyl-serine phosphorylation leukotriene biosynthetic process platelet activation neutrophil chemotaxis receptor internalization positive regulation of type I interferon production regulation of superoxide anion generation positive regulation of cell adhesion mediated by integrin peptidyl-tyrosine autophosphorylation Fc-epsilon receptor signaling pathway Fc-gamma receptor signaling pathway involved in phagocytosis defense response to bacterium transcription factor import into nucleus positive regulation of mast cell degranulation regulation of neutrophil degranulation beta selection innate immune response positive regulation of interleukin-3 biosynthetic process positive regulation of granulocyte macrophage colony-stimulating factor biosynthetic process positive regulation of B cell differentiation positive regulation of gamma-delta T cell differentiation positive regulation of bone resorption positive regulation of alpha-beta T cell differentiation positive regulation of alpha-beta T cell proliferation blood vessel morphogenesis positive regulation of cytokine secretion positive regulation of peptidyl-tyrosine phosphorylation regulation of phagocytosis positive regulation of calcium-mediated signaling B cell receptor signaling pathway regulation of sequence-specific DNA binding transcription factor activity regulation of ERK1 and ERK2 cascade cellular response to molecule of fungal origin cellular response to low-density lipoprotein particle stimulus regulation of arachidonic acid secretion regulation of platelet aggregation activation of MAPK activity serotonin secretion immune system process cell surface receptor signaling pathway enzyme linked receptor protein signaling pathway G-protein coupled receptor signaling pathway phosphorylation cell migration peptidyl-tyrosine phosphorylation signal transduction by protein phosphorylation cell differentiation intracellular signal transduction protein autophosphorylation regulation of immune response

参与通路

protein kinase activity protein serine/threonine kinase activity protein tyrosine kinase activity non-membrane spanning protein tyrosine kinase activity receptor signaling protein tyrosine kinase activity integrin binding protein binding ATP binding protein kinase binding nucleotide binding receptor binding kinase activity transferase activity transferase activity, transferring phosphorus-containing groups protein domain specific binding