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Invitrogen
Phospho-BTK/ITK (Tyr551, Tyr511) Monoclonal Antibody (M4G3LN), PE, eBioscience™
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FIGURE: 1 / 1
Phospho-BTK/ITK (Tyr551, Tyr511) Antibody (12-9015-41) in Flow
Intracellular staining of unstimulated (orange histogram) or 5-minute F (ab')2 Anti-Mouse IgM, u chain specific Functional Grade Purified-stimulated (Product # 16-5092-85) (purple histogram) mouse splenocytes with Anti-Human/Mouse phospho-BTK/ITK (Y551/Y511) PE. B220+ cells were used for analysis.
Product Details
12-9015-41
Applications
Tested Dilution
Publications
Product Specifications
Published species
Mouse
Host/Isotype
Mouse
/ IgG2b, kappa
Recommended Isotype Control
Class
Monoclonal
Type
Antibody
Clone
M4G3LN
Conjugate
Excitation/Emission Max
565/576 nm
View spectra
Form
Liquid
Concentration
5 µL/Test
Storage conditions
4°C, store in dark, DO NOT FREEZE!
Shipping conditions
Wet ice
RRID
AB_2572676
Product Specific Information
Description: 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 (1 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
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.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
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