Phospho-ZAP70/Syk (Tyr319, Tyr352) Monoclonal Antibody (n3kobu5), PE-Cyanine7, eBioscience
PRODUCT DETAILS
Host: Mouse
Isotype: IgG2b, kappa
Clonality: Monoclonal
Clone: n3kobu5
Format: PE-Cyanine7
Reactivity: Hu, Ms
Application: Flow Cytometry
Tested Dilution: 5 µL (0.06 µg)/test
Concentration: 5 μL/Test
Storage: 4°C, store in dark, DO NOT FREEZE!
Formulation: PBS with BSA and 0.09% sodium azide; pH 7.2
Purification: Affinity chromatography
Data Sheet: TDS
Specific Information
Description: This n3kobu5 monoclonal antibody recognizes human and mouse zeta chain-associated protein of 70 kD (also known as ZAP-70) and spleen tyrosine kinase (also known as SYK) when phosphorylated on Y319 and Y352, respectively. ZAP-70 and SYK are members of the SYK protein tyrosine kinase (PTK) family that are phosphorylated and activated by Src family PTKs. ZAP-70/SYK Y319/Y352 are located in the so-called interdomain of ZAP-70/SYK (between the N-terminal dual SH2 domains and the C-terminal kinase domain).
Phosphorylation of ZAP-70 Y319 by Lck is necessary for T cell receptor (TCR)-dependent association of ZAP-70 with Lck and phospholipase C gamma and subsequent activation of calcium-dependent and Ras signaling cascades. SYK Y352 phosphorylation by Fyn/Lyn is critical for propagation of B cell receptor (BCR) signaling and for B cell development.
Specificity of this n3kobu5 clone was determined by ELISA, flow cytometry, and western blotting.
Applications Reported: This n3kobu5 antibody has been reported for use in intracellular staining followed by flow cytometric analysis.
Applications Tested: This n3kobu5 antibody has been pre-diluted and tested by intracellular staining and flow cytometric analysis of sodium pervanadate-treated mouse splenocytes using the Intracellular Fixation & Permeabilization Buffer Set (a href="https://www.thermofisher.com/order/catalog/product/88-8824-00">Product # 88-8824-00) and protocol. Please refer to "Staining Intracellular Antigens for Flow Cytometry, Protocol A: Two step protocol for intracellular (cytoplasmic) proteins" located at www.thermofisher.com/flowprotocols . This may 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.
Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.
Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222-49) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333-57) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.
Excitation: 488-561 nm; Emission: 775 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Description
PRODUCT DETAILS
Host: Mouse
Isotype: IgG2b, kappa
Clonality: Monoclonal
Clone: n3kobu5
Format: PE-Cyanine7
Reactivity: Hu, Ms
Application: Flow Cytometry
Tested Dilution: 5 µL (0.06 µg)/test
Concentration: 5 μL/Test
Storage: 4°C, store in dark, DO NOT FREEZE!
Formulation: PBS with BSA and 0.09% sodium azide; pH 7.2
Purification: Affinity chromatography
Data Sheet: TDS
Specific Information
Description: This n3kobu5 monoclonal antibody recognizes human and mouse zeta chain-associated protein of 70 kD (also known as ZAP-70) and spleen tyrosine kinase (also known as SYK) when phosphorylated on Y319 and Y352, respectively. ZAP-70 and SYK are members of the SYK protein tyrosine kinase (PTK) family that are phosphorylated and activated by Src family PTKs. ZAP-70/SYK Y319/Y352 are located in the so-called interdomain of ZAP-70/SYK (between the N-terminal dual SH2 domains and the C-terminal kinase domain).
Phosphorylation of ZAP-70 Y319 by Lck is necessary for T cell receptor (TCR)-dependent association of ZAP-70 with Lck and phospholipase C gamma and subsequent activation of calcium-dependent and Ras signaling cascades. SYK Y352 phosphorylation by Fyn/Lyn is critical for propagation of B cell receptor (BCR) signaling and for B cell development.
Specificity of this n3kobu5 clone was determined by ELISA, flow cytometry, and western blotting.
Applications Reported: This n3kobu5 antibody has been reported for use in intracellular staining followed by flow cytometric analysis.
Applications Tested: This n3kobu5 antibody has been pre-diluted and tested by intracellular staining and flow cytometric analysis of sodium pervanadate-treated mouse splenocytes using the Intracellular Fixation & Permeabilization Buffer Set (a href="https://www.thermofisher.com/order/catalog/product/88-8824-00">Product # 88-8824-00) and protocol. Please refer to "Staining Intracellular Antigens for Flow Cytometry, Protocol A: Two step protocol for intracellular (cytoplasmic) proteins" located at www.thermofisher.com/flowprotocols . This may 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.
Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.
Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222-49) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333-57) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.
Excitation: 488-561 nm; Emission: 775 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
