Thermo Fisher Scientific Phospho-SRC (Tyr418) Monoclonal Antibody (SC1T2M3), PerCP-eFluor 710, eBioscience

Applications

Tested Dilution

Publications

Flow Cytometry (Flow)

5 µL (0.125 µg)/test

-

Product Specifications

Species Reactivity

Human, Mouse

Host/Isotype

Mouse / IgG2b, kappa

Recommended Isotype Control

Mouse IgG2b kappa Isotype Control (eBMG2b), PerCP-eFluor™ 710, eBioscience™

Class

Monoclonal

Type

Antibody

Clone

SC1T2M3

Conjugate

PerCP-eFluor™ 710 PerCP-eFluor™ 710 PerCP-eFluor™ 710

View additional formats

• Unconjugated
• eFluor 660
• PE
• Request custom conjugation

Excitation/Emission Max

482/708 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_2573866

Product Specific Information

Description: This SC1T2M3 monoclonal antibody recognizes human and mouse Src tyrosine kinase (also known as ASV, c-src, c-SRC, p60-Src, pp60c-src, Proto-oncogene c-Src, Proto-oncogene tyrosine-protein kinase Src, SRC1) when phosphorylated on tyrosine 418 (Y418). Autophosphorylation of Src at Y418 in the catalytic domain is required for full catalytic activity of this kinase. Src is a non-receptor tyrosine kinase involved in signal transduction in numerous biological systems and is activated following engagement of many different classes of cellular receptors including immune response receptors, integrins and other adhesion receptors, receptor protein tyrosine kinases, G protein-coupled receptors as well as cytokine receptors. Aberrant Src activity has been implicated in the development of numerous types of cancer. Due to the sequence homology surrounding Src Y418, this SC1T2M3 clone is predicted to cross-react with many Src family kinases including Src, Lck, Fyn, and Lyn.

Specificity of this SC1T2M3 clone was determined by ELISA, flow cytometry, and western blotting.

Applications Reported: This SC1T2M3 antibody has been reported for use in intracellular staining followed by flow cytometric analysis.

Applications Tested: This SC1T2M3 antibody has been pre-titrated and tested by intracellular staining followed by flow cytometric analysis of 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.

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.

PerCP-eFluor® 710 emits at 710 nm and is excited with the blue laser (488 nm); it can be used in place of PerCP-Cyanine5.5. We recommend using a 710/50 bandpass filter, however, the 695/40 bandpass filter is an acceptable alternative. Please make sure that your instrument is capable of detecting this fluorochrome.

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) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333) 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 nm; Emission: 710 nm; Laser: Blue Laser.

Filtration: 0.2 µm post-manufacturing filtered.

Target Information

SRC is a cytoplasmic tyrosine kinase and proto-oncogene involved in the regulation of embryonic development and cell growth. SRC is involved in the regulation of growth and differentiation of eukaryotic cells. SRC associates with the cytoplasmic region of various cell surface receptors to facilitate signal transduction, and is also involved in cytoskeletal organization, cell cycle regulation, and apoptosis. In humans, the gene encoding SRC is present on chromosome 20. Activity of SRC is regulated by two tyrosine residues (Tyr 416 and Tyr527), and phosphorylation of these two sites have opposing effects. Phosphorylation of Tyr416 in the activation loop of the kinase domain upregulates enzyme activity while the phosphorylation of Tyr527 decreases enzyme activity. Mutations in the SRC gene could be involved in the malignant progression of colon cancer. Two transcript variants encoding the same SRC protein have been found thus far.

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