SYK Monoclonal antibody

• Featured Product
• KD/KO Validated

SYK Monoclonal Antibody for FC, IF, IP, WB, ELISA

Host / Isotype

Mouse / IgG2b

Reactivity

Human and More (2)

Applications

WB, IP, IF, FC, ELISA

Conjugate

Unconjugated

Publications

2

CloneNo.

4C4A12

Cat no : 66721-1-Ig

Synonyms

DKFZp313N1010, FLJ25043, FLJ37489, spleen tyrosine kinase, SYK, Tyrosine protein kinase SYK

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Validation Data Gallery

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  WB analysis using 66721-1-Ig

  Various lysates were subjected to SDS PAGE followed by western blot with 66721-1-Ig (SYK antibody) at dilution of 1:5000 incubated at room temperature for 1.5 hours.

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  WB analysis of THP-1 using 66721-1-Ig

  THP-1 (WT and SYK KO) lysates prepared with RIPA buffer, 30 μg protein loaded. 66721-1-Ig incubated at 1:500 at 4°C overnight in 5% BSA in TBST. Ponceau stained transfers shown on right. Data provided by YCharOS, an open science company with a mission to validate commercial antibodies to improve scientific reproducibility and transparency.

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  WB analysis of Raji using 66721-1-Ig

  WB result of SYK antibody (66721-1-Ig; 1:10000; incubated at room temperature for 1.5 hours) with sh-Control and sh-SYK transfected Raji cells.

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  IP experiment of MDA-MB-231 using 66721-1-Ig

  MDA-MB-231 lysates prepared and IP of SYK performed using 2.0 μg of 66721-1-Ig coupled to protein G- Sepharose beads. The Ponceau stained transfers of each blot are shown. Data provided by YCharOS, an open science company with a mission to validate commercial antibodies to improve scientific reproducibility and transparency.

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  IF Staining of human tonsillitis using 66721-1-Ig

  Immunofluorescent analysis of (4% PFA) fixed human tonsillitis tissue using 66721-1-Ig (SYK antibody), at dilution of 1:100 and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L).

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  FC experiment of K-562 using 66721-1-Ig

  1X10^6 K-562 cells were intracellularly stained with 0.4 ug Anti-Human SYK (66721-1-Ig, Clone:4C4A12) and CoraLite®488-Conjugated AffiniPure Goat Anti-Mouse IgG(H+L) at dilution 1:1000 (red), or 0.4 ug Mouse IgG2b Isotype Control (MPC-11) (65128-1-Ig, Clone: MPC-11) (blue). Cells were fixed with 4% PFA and permeabilized with Flow Cytometry Perm Buffer (PF00011-C).

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Tested Applications

• Positive WB detected in: Raji cells, THP-1 cells, Ramos cells, Daudi cells
• Positive IP detected in: MDA-MB-231 cells
• Positive IF detected in: human tonsillitis tissue
• Positive FC detected in: K-562 cells

Recommended dilution

• Western Blot (WB): WB : 1:2000-1:10000
• Immunoprecipitation (IP): IP : 0.5-4.0 ug for 1.0-3.0 mg of total protein lysate
• Immunofluorescence (IF): IF : 1:50-1:500
• Flow Cytometry (FC): FC : 0.40 ug per 10^6 cells in a 100 µl suspension

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, Check data in validation data gallery.

Published Applications

• WB: See 2 publications below

Product Information

66721-1-Ig targets SYK in WB, IP, IF, FC, ELISA applications and shows reactivity with Human samples.

• Tested Reactivity: Human
• Cited Reactivity: human, mouse
• Host / Isotype: Mouse / IgG2b
• Class: Monoclonal
• Type: Antibody
• Immunogen: SYK fusion protein Ag17654
• Full Name: spleen tyrosine kinase
• Calculated Molecular Weight: 635 aa, 72 kDa
• Observed Molecular Weight: 70-75 kDa
• GenBank Accession Number: BC001645
• Gene Symbol: SYK
• Gene ID (NCBI): 6850
• RRID: AB_2882072
• Conjugate: Unconjugated
• Form: Liquid
• Purification Method: Protein A purification
• Storage Buffer: PBS with 0.02% sodium azide and 50% glycerol pH 7.3.
• Storage Conditions: Store at -20°C. Stable for one year after shipment. Aliquoting is unnecessary for -20^oC storage. 20ul sizes contain 0.1% BSA.

Background Information

SYK(spleen tyrosine kinase) is a key regulator of signal transduction events,apoptosis and orderly cell cycle progression in B-lineage lymphoid cells. Although SYK has not been linked to a human disease, defective expression of the closely related T-cell tyrosine kinase ZAP-70 has been associated with severe combined immunodeficiency(PMID:11494125). This protein has 2 isoforms produced by alternative splicing. SYK also promotes liver fibrosis by activation of hepatic stellate cells and acts as a biomarker for human hepatocellular carcinoma. (PMID: 29537660)

Publications

Species	Application	Title
mouse	WB	Cancer Lett; SIGLEC15 amplifies immunosuppressive properties of tumor-associated macrophages in pancreatic cancer.; Authors - Tian-Jiao Li
human	WB	Exp Cell Res; CD93 promotes acute myeloid leukemia development and is a potential therapeutic target; Authors - Jie Jia