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Invitrogen

Phospho-BTK/ITK (Tyr551, Tyr511) Monoclonal Antibody (M4G3LN), APC, eBioscience™

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Product Details

17-9015-41

Applications
Tested Dilution
Publications

Flow Cytometry (Flow)

5 µL (0.125 µg)/test
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Product Specifications

Species Reactivity

Human, Mouse

Published species

Mouse

Host/Isotype

Mouse / IgG2b, kappa

Recommended Isotype Control

Mouse IgG2b kappa Isotype Control (eBMG2b), APC, eBioscience™

Class

Monoclonal

Type

Antibody

Clone

M4G3LN

Conjugate

APC APC APC

Excitation/Emission Max

651/660 nm View spectra spectra

Form

Liquid

Concentration

5 µL/Test

Purification

Affinity chromatography

Storage buffer

PBS, pH 7.2, with 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_2573277

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 (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.

Excitation: 633-647 nm; Emission: 660 nm; Laser: Red 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.

How to use the Panel Builder

Watch the video to learn how to use the Invitrogen Flow Cytometry Panel Builder to build your next flow cytometry panel in 5 easy steps.

Bioinformatics

Protein Aliases: agammaglobulinaemia tyrosine kinase; Agammaglobulinemia tyrosine kinase; ATK; B-cell progenitor kinase; BPK; Bruton agammaglobulinemia tyrosine kinase; Bruton tyrosine kinase; Bruton's tyrosine kinase; dominant-negative kinase-deficient Brutons tyrosine kinase; frameshift mutation results in truncated protein; homolog of mouse T-cell itk/tsk; IL-2-inducible T-cell kinase; interleukin-2-inducible T cell kinase; Interleukin-2-inducible T-cell kinase; Kinase EMB; Kinase EMT; Kinase TLK; T-cell-specific kinase; Tyrosine-protein kinase BTK; Tyrosine-protein kinase BTK (Brutons tyrosine kinase) (Agammaglobulinaemia tyrosine kinase) (ATK) (B cell progenitor kinase) (BPK) (Kinase EMB); Tyrosine-protein kinase ITK/TSK; Tyrosine-protein kinase Lyk; unnamed protein product; X-linked immune deficiency

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Gene Aliases: AGMX1; AI528679; AT; ATK; BPK; BTK; EMT; IGHD3; IMD1; ITK; LPFS1; LYK; PSCTK1; PSCTK2; Tcsk; Tlk; Tsk; xid; XLA

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UniProt ID: (Human) Q06187, (Mouse) P35991, (Human) Q08881, (Mouse) Q03526

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Entrez Gene ID: (Human) 695, (Mouse) 12229, (Human) 3702, (Mouse) 16428

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Function(s)
nucleotide binding protein kinase activity protein tyrosine kinase activity non-membrane spanning protein tyrosine kinase activity protein binding ATP binding phosphatidylinositol-3,4,5-trisphosphate binding zinc ion binding lipid binding phospholipase activator activity kinase activity transferase activity identical protein binding phospholipase binding metal ion binding transferase activity, transferring phosphorus-containing groups GTPase-activating protein
Process(es)
neutrophil homeostasis positive regulation of type III hypersensitivity positive regulation of type I hypersensitivity negative regulation of cytokine production adaptive immune response B cell affinity maturation immune system process histamine secretion by mast cell positive regulation of immunoglobulin production regulation of B cell cytokine production MyD88-dependent toll-like receptor signaling pathway regulation of B cell apoptotic process apoptotic process mesoderm development peptidyl-tyrosine phosphorylation calcium-mediated signaling proteoglycan catabolic process negative regulation of B cell proliferation positive regulation of B cell proliferation positive regulation of type I interferon production response to lipopolysaccharide negative regulation of interleukin-10 production positive regulation of interleukin-6 production positive regulation of tumor necrosis factor production cellular response to reactive oxygen species intracellular signal transduction Fc-epsilon receptor signaling pathway B cell activation positive regulation of I-kappaB kinase/NF-kappaB signaling innate immune response positive regulation of B cell differentiation cell maturation positive regulation of phagocytosis T cell receptor signaling pathway B cell receptor signaling pathway regulation of B cell activation negative regulation of B cell activation positive regulation of NF-kappaB transcription factor activity monocyte proliferation negative regulation of leukocyte proliferation cellular response to molecule of fungal origin apoptotic signaling pathway cellular response to interleukin-7 cGAS/STING signaling pathway positive regulation of cGAS/STING signaling pathway positive regulation of interleukin-17A production positive regulation of NLRP3 inflammasome complex assembly positive regulation of synoviocyte proliferation eosinophil homeostasis transcription, DNA-templated regulation of transcription, DNA-templated protein phosphorylation transmembrane receptor protein tyrosine kinase signaling pathway I-kappaB kinase/NF-kappaB signaling response to organic substance phosphorylation cell differentiation peptidyl-tyrosine autophosphorylation regulation of cell proliferation protein autophosphorylation positive regulation of cytokine production NK T cell differentiation cellular defense response signal transduction T cell activation gamma-delta T cell activation cytokine production activation of phospholipase C activity interferon-gamma production interleukin-4 production
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