Phospho-BTK/ITK (Tyr551, Tyr511), PerCP-eFluor™ 710, clone: M4G3LN, eBioscience™
Mouse Monoclonal Antibody
Brand: Affymetrix eBioscience 46-9015-41
Code : NEW
Additional Details : Weight : 0.23750kg
DescriptionDescription: This M4G3LN monoclonal antibody recognizes human and mouse Bruton's tyrosine kinase (also known as BTK, agammaglobulinaemia tyrosine kinase, B-cell progenitor kinase)/interleukin-2-inducible tyrosine kinase (also known as ITK, T-cell-specific kinase, Tyrosine-protein kinase Lyk) when phosphorylated on Y551 and Y511, respectively.In addition to Src family and SYK family kinases, B cells and T cells express a third family of receptor-proximal kinases called the Tec kinase family. The B cell-specific Tec family kinase is BTK; T cells express ITK as well as Tec and Rlk. Tec family kinases play an indispensable role in activation of phospholipase C and activation of calcium-dependent signaling cascades following antigen receptor triggering. BTK has been implicated in propagating signals downstream of LPS-TLR4 in B cells via signaling through the TLR4 co-receptor RP105/CD180. Mice congenitally lacking BTK or ITK show profound defects in B cell or T cell development and signaling, respectively. People with mutations or deletions in BTK suffer from X-linked agammaglobulinemia and X-linked immunodeficiency.Specificity of this M4G3LN clone was determined by ELISA, flow cytometry, and western blotting.Applications Reported: This M4G3LN antibody has been reported for use in intracellular staining followed by flow cytometric analysis.Applications Tested: This M4G3LN antibody has been pre-titrated and tested by intracellular staining followed by flow cytometric analysis of stimulated normal human peripheral blood cells. This can be used at 5 μL (0.5 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 Best Protocols 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 Best Protocols 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.Fixation: Samples can be stored in IC Fixation Buffer (cat. 00-8222) (100 μL of cell sample + 100 μL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (cat. 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: 0.2μm post-manufacturing filtered.
|PBS with 0.1% gelatin, 0.2% BSA and 0.09% sodium azide; pH 7.2|
|Q06187, Q08881, Q03526, P35991|
|agammaglobulinaemia tyrosine kinase, AGMX1, ATK, B-cell progenitor kinase, BPK, Bruton agammaglobulinemia tyrosine kinase, Bruton's tyrosine kinase, dominant-negative kinase-deficient Brutons tyrosine kinase, truncated Bruton agammaglobulinemia tyrosine kinase, tyrosine-protein kinase BTK isoform (lacking exon 13 to 17), tyrosine-protein kinase BTK isoform (lacking exon 14), EMT, LYK, PSCTK2, IL-2-inducible T cell kinase, homolog of mouse T-cell itk/tsk, IL-2-inducible T cell kinase, IL-2-inducible T-cell kinase, IL2-inducible T-cell kinase, interleukin-2-inducible T cell kinase, interleukin-2-inducible T-cell kinase, kinase EMT, T-cell-specific kinase, tyrosine-protein kinase ITK/TSK, tyrosine-protein kinase LYK|
|Store at 2-8°C. Do not freeze. Light-sensitive material. This tandem dye is sensitive to photo-induced oxidation. Protect this vial from light during storage, handling and experimental procedures.|
For Research Use Only.