BTK [p Tyr551] Antibody (797837) [Unconjugated]

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.

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.

Antigen-triggered phosphorylation of BTK and PLC gamma in BMMCs is independent of PI3K. (a) WT BMMCs were pre-treated for 20 min with 100 nM Wortmannin (WM) or vehicle (DMSO) and were then left untreated (con) or stimulated with the indicated concentrations of Ag (DNP-HSA [DNP]) for 1 and 5 min. Btk−/− BMMCs were pre-treated with DMSO and stimulated accordingly. Whole-cell lysates were subjected to Western Blot analysis with antibodies against P-BTK (Tyr223), P-PKB (Ser473), and p85 (loading control). (b) WT BMMCs were pre-treated for 20 min with 100 nM WM or vehicle (DMSO) and were then left untreated (con) or stimulated with Ag (20 ng/ml DNP-HSA) for the indicated times. Btk−/− BMMCs were pre-treated with DMSO and stimulated accordingly. Whole-cell lysates were subjected to Western Blot analysis with antibodies against P-BTK (Tyr551), BTK, P-PKB (Ser473), and p85 (loading control). (c) WT and Btk−/− BMMCs were pre-treated for 20 min with 100 nM WM or vehicle (DMSO) and were then left untreated (con) or stimulated with the indicated concentrations of Ag for 1 and 5 min. Whole-cell lysates were analysed by immunoblotting against P-PLC gamma 1 (Tyr783), PLC gamma 1, P-PKB (Ser473), and p85 (loading control). Densitometry of P-PLC gamma 1 with reference to PLC gamma 1 was performed using ImageJ software (NIH, USA) and normalised to WT cells pretreated with DMSO and stimulated with 1 ng/ml Ag (1 min). Comparable results were obtained in at least three experiments with different BMMC cultures. Fine vertical lines were inserted to allow for better discrimination of cells/conditions. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/30341350), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: WB, Simple Western
• Clone: 797837
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: LYOPH

BTK [p Tyr551] Antibody (797837) [Unconjugated] Summary

• Immunogen: Phosphopeptide containing the human BTK Y551 site
• Modification: p Tyr551
• Specificity: Detects human BTK when phosphorylated at Y551.
• Source: N/A
• Isotype: IgG2a
• Clonality: Monoclonal
• Host: Mouse
• Gene: BTK
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Simple Western 20 ug/mL
  • Western Blot 1 ug/mL
• Application Notes: In Simple Western only 10-15 uL of the recommended dilution is used per data point.

Packaging, Storage & Formulations

• 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.
• Buffer: 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.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Sterile PBS to a final concentration of 0.5 mg/mL.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for BTK [p Tyr551] Antibody (797837) [Unconjugated]

• 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

Background

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.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for BTK Antibody (MAB7659)(2)

Publications using MAB7659

• CN Zorn, A Simonowski, M Huber Stimulus strength determines the BTK-dependence of the SHIP1-deficient phenotype in IgE/antigen-triggered mast cells Sci Rep, 2018-10-19;8(1):15467. 2018-10-19 [PMID: 30341350] (Western Blot, Mouse)
• SD Reiff, R Mantel, LL Smith, JT Greene, EM Muhowski, CA Fabian, VM Goettl, M Tran, B Harrington, KA Rogers, FT Awan, K Maddocks, L Andritsos, AM Lehman, D Sampath, R Lapalombel, S Eathiraj, G Abbadessa, B Schwartz, A Johnson, JC Byrd, JA Woyach The BTK Inhibitor ARQ 531 Targets Ibrutinib Resistant CLL and Richter's Transformation Cancer Discov, 2018-08-09;0(0):. 2018-08-09 [PMID: 30093506] (Western Blot, Human)

Bioinformatics

• Gene Symbol: BTK