Thermo Fisher Scientific Phospho-Syk (Tyr348) Monoclonal Antibody (moch1ct), PE, eBioscience

Applications

Tested Dilution

Publications

Flow Cytometry (Flow)

5 µL (0.06 µg)/test

View 2 publications 2 publications

Product Specifications

Species Reactivity

Human, Mouse

Published species

Mouse

Host/Isotype

Mouse / IgG1, kappa

Recommended Isotype Control

Mouse IgG1 kappa Isotype Control (P3.6.2.8.1), PE, eBioscience™

Class

Monoclonal

Type

Antibody

Clone

moch1ct

Conjugate

PE PE PE

View additional formats

• APC
• Request custom conjugation

Excitation/Emission Max

565/576 nm View spectra

Form

Liquid

Concentration

5 µL/Test

Purification

Affinity chromatography

Storage buffer

PBS, pH 7.2, with 0.2% BSA

Contains

0.09% sodium azide

Storage conditions

4° C, store in dark, DO NOT FREEZE!

Shipping conditions

Ambient (domestic); Wet ice (international)

RRID

AB_2572674

Product Specific Information

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 B cell 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 BestProtocols 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 BestProtocols® 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.

Target Information

Spleen tyrosine kinase (SYK), along with ZAP70, are cytoplasmic tyrosine kinases that are viewed as potential druge targets for allergic disorders and autoimmune diseases. SYK may also play a role in cancer. Syk 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.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.