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  • Product Name
    Phospho-BTK (Tyr223) Antibody
  • Catalog No.
  • RRID
  • Source
  • Application
  • Reactivity
    Human, Mouse, Rat
  • Prediction
    Pig(100%), Bovine(100%), Horse(100%), Sheep(100%), Rabbit(100%), Dog(100%)
  • UniProt
  • Mol.Wt
  • Concentration
  • Browse similar products>>

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Product Information

Alternative Names:Expand▼

Agammaglobulinaemia tyrosine kinase; AGMX 1; AGMX1; AT; ATK; B cell progenitor kinase; B-cell progenitor kinase; BPK; Bruton agammaglobulinemia tyrosine kinase; Bruton tyrosine kinase; Bruton’s Tyrosine Kinase; Btk; BTK_HUMAN; dominant-negative kinase-deficient Brutons tyrosine kinase; IMD 1; IMD1; MGC126261; MGC126262; OTTHUMP00000063593; PSCTK 1; PSCTK1; truncated Bruton agammaglobulinemia tyrosine kinase; Tyrosine protein kinase BTK; Tyrosine-protein kinase BTK; tyrosine-protein kinase BTK isoform (lacking exon 14; XLA;


WB 1:500-1:2000, IF/ICC 1:100-1:500, ELISA(peptide) 1:20000-1:40000
*The optimal dilutions should be determined by the end user.


Human, Mouse, Rat

Predicted Reactivity:

Pig(100%), Bovine(100%), Horse(100%), Sheep(100%), Rabbit(100%), Dog(100%)






The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.


Phospho-BTK (Tyr223) Antibody detects endogenous levels of BTK only when phosphorylated at Tyrosine 223.


Please cite this product as: Affinity Biosciences Cat# AF0841, RRID:AB_2834080.





Storage Condition and Buffer:

Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol.Store at -20 °C.Stable for 12 months from date of receipt.

Immunogen Information


A synthesized peptide derived from human BTK around the phosphorylation site of Tyr223.


>>Visit The Human Protein Atlas

Gene ID:

Gene Name:


Molecular Weight:

Observed Mol.Wt.: 80kD.
Predicted Mol.Wt.: 76kDa(Calculated)..

Subcellular Location:

Cytoplasm. Membrane. Nucleus.

Tissue Specificity:

Predominantly expressed in B-lymphocytes.


Defects in the Bruton tyrosine kinase (BTK) gene cause Agammaglobulinemia. Agammaglobulinemia is an X-linked immunodeficiency characterized by failure to produce mature B lymphocyte cells and associated with a failure of Ig heavy chain rearrangement. [provided by RefSeq]


Research Background


Non-receptor tyrosine kinase indispensable for B lymphocyte development, differentiation and signaling. Binding of antigen to the B-cell antigen receptor (BCR) triggers signaling that ultimately leads to B-cell activation. After BCR engagement and activation at the plasma membrane, phosphorylates PLCG2 at several sites, igniting the downstream signaling pathway through calcium mobilization, followed by activation of the protein kinase C (PKC) family members. PLCG2 phosphorylation is performed in close cooperation with the adapter protein B-cell linker protein BLNK. BTK acts as a platform to bring together a diverse array of signaling proteins and is implicated in cytokine receptor signaling pathways. Plays an important role in the function of immune cells of innate as well as adaptive immunity, as a component of the Toll-like receptors (TLR) pathway. The TLR pathway acts as a primary surveillance system for the detection of pathogens and are crucial to the activation of host defense. Especially, is a critical molecule in regulating TLR9 activation in splenic B-cells. Within the TLR pathway, induces tyrosine phosphorylation of TIRAP which leads to TIRAP degradation. BTK plays also a critical role in transcription regulation. Induces the activity of NF-kappa-B, which is involved in regulating the expression of hundreds of genes. BTK is involved on the signaling pathway linking TLR8 and TLR9 to NF-kappa-B. Transiently phosphorylates transcription factor GTF2I on tyrosine residues in response to BCR. GTF2I then translocates to the nucleus to bind regulatory enhancer elements to modulate gene expression. ARID3A and NFAT are other transcriptional target of BTK. BTK is required for the formation of functional ARID3A DNA-binding complexes. There is however no evidence that BTK itself binds directly to DNA. BTK has a dual role in the regulation of apoptosis.

Post-translational Modifications:

Following B-cell receptor (BCR) engagement, translocates to the plasma membrane where it gets phosphorylated at Tyr-551 by LYN and SYK. Phosphorylation at Tyr-551 is followed by autophosphorylation of Tyr-223 which may create a docking site for a SH2 containing protein. Phosphorylation at Ser-180 by PRKCB, leads in translocation of BTK back to the cytoplasmic fraction. Phosphorylation at Ser-21 and Ser-115 creates a binding site for PIN1 at these Ser-Pro motifs, and promotes it's recruitment.

Subcellular Location:

Cytoplasm. Cell membrane>Peripheral membrane protein. Nucleus.
Note: In steady state, BTK is predominantly cytosolic. Following B-cell receptor (BCR) engagement by antigen, translocates to the plasma membrane through its PH domain. Plasma membrane localization is a critical step in the activation of BTK. A fraction of BTK also shuttles between the nucleus and the cytoplasm, and nuclear export is mediated by the nuclear export receptor CRM1.

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionGraphics by Christian Stolte

Tissue Specificity:

Predominantly expressed in B-lymphocytes.

Subunit Structure:

Binds GTF2I through the PH domain. Interacts with SH3BP5 via the SH3 domain. Interacts with IBTK via its PH domain. Interacts with ARID3A, CAV1, FASLG, PIN1, TLR8 and TLR9.


The PH domain mediates the binding to inositol polyphosphate and phosphoinositides, leading to its targeting to the plasma membrane. It is extended in the BTK kinase family by a region designated the TH (Tec homology) domain, which consists of about 80 residues preceding the SH3 domain.

Belongs to the protein kinase superfamily. Tyr protein kinase family. TEC subfamily.

Research Fields

Research Fields:

· Environmental Information Processing > Signal transduction > NF-kappa B signaling pathway.(View pathway)
· Human Diseases > Immune diseases > Primary immunodeficiency.
· Organismal Systems > Immune system > Platelet activation.(View pathway)
· Organismal Systems > Immune system > Fc epsilon RI signaling pathway.(View pathway)
· Organismal Systems > Development > Osteoclast differentiation.(View pathway)
· Organismal Systems > Immune system > B cell receptor signaling pathway.(View pathway)

WB Images(1)

IF/ICC Images(1)

ELISA analysis of AF0841 showing specificity to Phospho-BTK (Tyr223) Blocking Peptide peptide. Peptides concentration: 1ug/ml.
P-peptide: phospho-peptide; N-peptide: non-phospho-peptide.
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Catalog Number :

(Blocking peptide available as AF0841-BP)

Price/Size :

Tips: For phospho antibody, we provide phospho peptide(0.5mg) and non-phospho peptide(0.5mg).

Function :

Blocking peptides are peptides that bind specifically to the target antibody and block antibody binding. These peptide usually contains the epitope recognized by the antibody. Antibodies bound to the blocking peptide no longer bind to the epitope on the target protein. This mechanism is useful when non-specific binding is an issue, for example, in Western blotting (immunoblot) and immunohistochemistry (IHC). By comparing the staining from the blocked antibody versus the antibody alone, one can see which staining is specific; Specific binding will be absent from the western blot or immunostaining performed with the neutralized antibody.

Format and storage :

Synthetic peptide was lyophilized with 100% acetonitrile and is supplied as a powder. Reconstitute with 0.1 ml DI water for a final concentration of 10 mg/ml.The purity is >90%,tested by HPLC and MS.Storage Maintain refrigerated at 2-8°C for up to 6 months. For long term storage store at -20°C.

Precautions :

This product is for research use only. Not for use in diagnostic or therapeutic procedures.

High similarity Medium similarity Low similarity No similarity
Q06187 as Substrate
Site PTM Type Enzyme
A2 Acetylation
S21 Phosphorylation
Y40 Phosphorylation
S51 Phosphorylation P31749 (AKT1)
S55 Phosphorylation
S93 Phosphorylation
S115 Phosphorylation
S179 Phosphorylation
S180 Phosphorylation P05771 (PRKCB)
K183 Acetylation
T184 Phosphorylation
T191 Phosphorylation
Y223 Phosphorylation Q06187 (BTK) , Q08881 (ITK) , P07948 (LYN) , P00519 (ABL1) , A0A173G4P4 (Abl fusion) , P42680 (TEC)
Y225 Phosphorylation
Y263 Phosphorylation
Y279 Phosphorylation
K300 Ubiquitination
S318 Phosphorylation
K322 Ubiquitination
Y334 Phosphorylation
S342 Phosphorylation
Y344 Phosphorylation
Y345 Phosphorylation
Y361 Phosphorylation
S366 Phosphorylation
S371 Phosphorylation
Y375 Phosphorylation
S378 Phosphorylation
K406 Ubiquitination
Y461 Phosphorylation
K466 Ubiquitination
T495 Phosphorylation P31749 (AKT1)
K536 Ubiquitination
S543 Phosphorylation
Y551 Phosphorylation P07948 (LYN) , P12931 (SRC) , Q06187 (BTK) , P43405 (SYK)
T552 Phosphorylation
S553 Phosphorylation
K558 Ubiquitination
S564 Phosphorylation
T602 Phosphorylation
S604 Phosphorylation
Y617 Phosphorylation
S623 Phosphorylation
S659 Phosphorylation
Q06187 as PTM Enzyme
Substrate Site Source
P15391-1 (CD19) Y500 Uniprot
P15391-1 (CD19) Y531 Uniprot
P16220 (CREB1) S133 Uniprot
P16885 (PLCG2) Y753 Uniprot
P16885 (PLCG2) Y759 Uniprot
P16885 (PLCG2) Y1197 Uniprot
P16885 (PLCG2) Y1217 Uniprot
P42229 (STAT5A) Y694 Uniprot
P42680 (TEC) Y206 Uniprot
P42768 (WAS) Y291 Uniprot
P51813 (BMX) Y216 Uniprot
P51813 (BMX) Y224 Uniprot
P58753 (TIRAP) Y86 Uniprot
P58753 (TIRAP) Y106 Uniprot
P58753 (TIRAP) Y187 Uniprot
P78347-2 (GTF2I) Y248 Uniprot
P78347-2 (GTF2I) Y357 Uniprot
P78347-4 (GTF2I) Y377 Uniprot
P78347-3 (GTF2I) Y378 Uniprot
P78347 (GTF2I) Y398 Uniprot
P78347-2 (GTF2I) Y462 Uniprot
P78347-4 (GTF2I) Y482 Uniprot
P78347-3 (GTF2I) Y483 Uniprot
P78347 (GTF2I) Y503 Uniprot
Q06187 (BTK) Y223 Uniprot
Q06187 (BTK) Y551 Uniprot
Q08881 (ITK) Y180 Uniprot
Q13422 (IKZF1) S214 Uniprot
Q13422 (IKZF1) S215 Uniprot
Q9UJV9 (DDX41) Y364 Uniprot
Q9UJV9 (DDX41) Y414 Uniprot
Q9UN19-1 (DAPP1) Y139 Uniprot
IMPORTANT: For western blots, incubate membrane with diluted antibody in 5% w/v milk , 1X TBS, 0.1% Tween®20 at 4°C with gentle shaking, overnight.

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