Human Phospho-BTK (Y223) Antibody

Name: Human Phospho-BTK (Y223) Antibody
Brand: R&D Systems
SKU: MAB7205
Rating: 5 (2 reviews)

Catalog #: MAB7205 2 Reviews Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
MAB7205
MAB7205-SP

Detection of Human Phospho-BTK (Y223) by Western Blot.

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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human Phospho-BTK (Y223) Antibody Summary

Species Reactivity

Human

Specificity

Detects human Phospho-BTK (Y223) in direct ELISAs and Western blots.

Source

Monoclonal Mouse IgG_2B Clone # 720101

Purification

Protein A or G purified from hybridoma culture supernatant

Immunogen

Phosphopeptide containing the human BTK Y223 site

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Applications

Recommended Concentration

Sample

Western Blot

1 µg/mL

See below

Immunocytochemistry

8-25 µg/mL

See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Western Blot

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Detection of Human Phospho-BTK (Y223) by Western Blot. Western blot shows lysates of Raji human Burkitt's lymphoma cell line untreated (-) or treated (+) with 1 mM Pervanadate for 30 minutes. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human Phospho-BTK (Y223) Monoclonal Antibody (Catalog # MAB7205) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Phospho-BTK (Y223) at approximately 80 kDa (as indicated). For additional reference, duplicate samples were probed with 0.05 µg/mL of Mouse Anti-Human BTK Monoclonal Antibody (lower panel, Catalog # MAB5807) This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Immunocytochemistry

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Phospho-BTK (Y223) in Raji Human Cell Line. BTK phosphorylated at Y223 was detected in immersion fixed Raji human Burkitt's lymphoma cell line with (upper panel) and without (lower panel) pervanadate stimulation using Mouse Anti-Human Phospho-BTK (Y223) Monoclonal Antibody (Catalog # MAB7205) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; Catalog # NL007) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Sterile PBS to a final concentration of 0.5 mg/mL.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: BTK

Bruton tyrosine kinase (BTK) is a 75 kDa cytoplasmic protein tyrosine kinase that is widely expressed in hematopoietic cells. BTK is required for B cell receptor signaling and B cell development. Defects in BTK result in X-linked agammaglobulinemia which is characterized by a severely decreased level of circulating antibodies. Like other Tec family kinases, BTK contains a Pleckstrin homology domain, a Tec homology domain, an SH3 domain, an SH2 domain, and a protein kinase domain. Association of the BTK SH2 domain with the B cell linker protein (BLNK) is required for the activation of PLC gamma by BTK. The autophosphorylation of Tyr223 facilitates the interaction of BTK with activated Syk. The sequence used as a peptide immunogen is identical in human, mouse and rat BTK.

Long Name

Bruton Agammaglobulinemia Tyrosine Kinase

Entrez Gene IDs

695 (Human)

Alternate Names

Agammaglobulinaemia tyrosine kinase; AGMX1; AGMX1MGC126262; AT; ATKIMD1; B-cell progenitor kinase; BPK; Bruton agammaglobulinemia tyrosine kinase; Bruton tyrosine kinase; BTK; dominant-negative kinase-deficient Brutons tyrosine kinase; EC 2.7.10; EC 2.7.10.2; IMD1; PSCTK1; tyrosine-protein kinase BTK; XLA; XLAMGC126261