Detection of Human Phospho-BTK (Y551) by Western Blot.
Western blot shows lysates of RL human non-Hodgkin's lymphoma B cell line and Ramos human Burkitt's lymphoma cell line untreated (-) or treated (+) with 0.1 mM Pervanadate (PV) for 10 minutes and 12 µg/mL Goat Anti-Human IgM μ Chain Antigen Affinity-purified Polyclonal Antibody (Catalog # G‑105‑C) for 2 minutes. PVDF membrane was probed with 1 µg/mL of Mouse Anti-Human Phospho-BTK (Y551) Monoclonal Antibody (Catalog # MAB7659) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). A specific band was detected for Phospho-BTK (Y551) at approximately 75 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Detection of Human Phospho-BTK (Y551) by Simple WesternTM.
Simple Western lane view shows lysates of Ramos human Burkitt's lymphoma cell line untreated (-) or treated (+) with ultraviolet light (UV), loaded at 0.2 mg/mL. A specific band was detected for Phospho-BTK (Y551) at approximately 89 kDa (as indicated) using 20 µg/mL of Mouse Anti-Human Phospho-BTK (Y551) Monoclonal Antibody (Catalog # MAB7659). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Preparation and Storage
Sterile PBS to a final concentration of 0.5 mg/mL.
Reconstitution Buffer Available
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
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.
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 PLCg by
Btk. Btk can be activated by Lyn or Syk phosphorylation of Tyr551 within the
catalytic domain. Within aa 546 - 556, human Btk shares 100% aa sequence
identity with mouse and rat Btk.
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