PRODUCT DETAILS

Host: Mouse

Isotype: IgG2b, kappa

Clonality: Monoclonal

Clone: pY20

Format: eFluor 450

Reactivity: Chem

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: The pY20 monoclonal antibody recognizes phosphorylated tyrosine residues (p-Tyr) on proteins. Numerous intracellular signaling cascades are propagated via phosphorylation of specific tyrosine on signaling proteins. The detection of p-Tyr residues is valuable for the characterization and purification of phosphorylated proteins and the biochemical pathways that they are involved in.

Applications Reported:This pY20 antibody has been reported for use in intracellular staining followed by flow cytometric analysis.

Applications Tested: This pY20 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 (0.06 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.

eFluor™ 450 is an alternative to Pacific Blue™. eFluor™ 450 emits at 445 nm and is excited with the Violet laser (405 nm). Please make sure that your instrument is capable of detecting this fluorochrome.

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) protein(s). 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: 405 nm; Emission: 445 nm; Laser: Violet Laser.

Filtration: 0.2 µm post-manufacturing filtered.

 

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