Revision 1

#72159Store at -20C

1 Kit

(9 x 20 microliters)

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
CD79A (D1X5C) XP® Rabbit mAb 13333 20 µl 45-55 kDa Rabbit IgG
Phospho-CD79A (Tyr182) Antibody 5173 20 µl 45-55 kDa Rabbit 
CD19 (Intracellular Domain) (D4V4B) XP® Rabbit mAb 90176 20 µl 95 kDa Rabbit IgG
Phospho-CD19 (Tyr531) Antibody 3571 20 µl 95 kDa Rabbit 
Phospho-Btk (Tyr223) (D1D2Z) Rabbit mAb 87457 20 µl 78 kDa Rabbit IgG
Syk (D3Z1E) XP® Rabbit mAb 13198 20 µl 72 kDa Rabbit IgG
Phospho-Syk (Tyr525/526) (C87C1) Rabbit mAb 2710 20 µl 72 kDa Rabbit IgG
Phospho-BLNK (Tyr96) Antibody 3601 20 µl 68, 70 kDa Rabbit 
Btk (D3H5) Rabbit mAb 8547 20 µl 77 kDa Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

The B Cell Signaling Antibody Sampler Kit II provides an economical means to examine key signaling proteins commonly associated with B cell activation. The provided antibodies allow monitoring of both total protein levels and the phosphorylation state. The kit includes enough antibody to perform two western blot experiments with each primary antibody.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Background

Antigen receptors found on the surface of B cells contain a heterodimeric signaling component composed of CD79A and CD79B, also known as Ig α and Ig ß, respectively. Presence of this receptor complex is essential for B-cell development and function. Antigen binding precedes formation of the CD79A and CD79B heterodimer and subsequent activation of receptor associated kinases. Tyr182 of mouse CD79A (corresponding to Tyr188 of human CD79A) is one of two key tyrosine residues in the immunoreceptor tyrosine-based activation motif (ITAM) of CD79A that are phosphorylated by Src family kinases (e.g., Lyn, Blk), and play a critical role in modulating signal transduction following immune receptor activation.

Syk is a protein tyrosine kinase that plays an important role in intracellular signal transduction in hematopoietic cells (1-3). Syk interacts with immunoreceptor tyrosine-based activation motifs (ITAMs) located in the cytoplasmic domains of immune receptors (4). It couples the activated immunoreceptors to downstream signaling events that mediate diverse cellular responses, including proliferation, differentiation, and phagocytosis (4). There is also evidence that Syk plays a role in nonimmune cells; Syk is a potential tumor suppressor in human breast carcinomas (5). Tyrosine 525 and 526 are located in the activation loop of the Syk kinase domain, and phosphorylation of Tyr525/526 of human Syk (equivalent to the Tyr519/520 of mouse Syk) is essential for Syk function (6).

Lyn, one of the Src family members, is predominantly expressed in hematopoietic cells (7). Two tyrosine residues have been reported to play a crucial role in the regulation of protein tyrosine kinases of the Src family. Autophosphorylation of Tyr396 (equivalent to Tyr416 of Src), located in the catalytic domain, correlates with enzyme activation. Csk-mediated phosphorylation of the carboxy-terminal Tyr507 (equivalent to Tyr527 of Src) inactivates the kinase. Tyrosine phosphorylation and activation of Lyn occurs upon association with cell surface receptors such as the B cell Ag receptor (BCR) and CD40 (8-10).

Bruton's tyrosine kinase (Btk) is a member of the Btk/Tec family of cytoplasmic tyrosine kinases. Btk plays an important role in B cell development (11,12). Activation of B cells by various ligands is accompanied by Btk membrane translocation mediated by its PH domain binding to phosphatidylinositol-3,4,5-trisphosphate (13-15). The membrane-located Btk is active and associated with transient phosphorylation of two tyrosine residues, Tyr551 and Tyr223. Tyr551 in the activation loop is transphosphorylated by the Src family tyrosine kinase, leading to autophosphorylation at Tyr223 within the SH3 domain, which is necessary for full activation (16,17).

CD19 is a 95 kDa coreceptor that amplifies the signaling cascade in B cells (18). On the B cell surface, CD19 associates with CD21, CD81, and Leu-13 to exert its function. The cytoplasmic tail of CD19 has nine conserved tyrosine residues playing critical roles in CD19-mediated function by coupling signaling molecules to the receptor (18). After BCR or CD19 ligation, Tyr531 and Tyr500 of CD19 are progressively phosphorylated. This phosphorylation enables the coupling of PI3 kinase and Src family tyrosine kinase to CD19 and activates the PI3K and Src signaling pathways (19,20).

B cell linker protein (BLNK), also known as SLP-65 or BASH, is an adaptor molecule that plays key roles in B cell activation and B cell antigen receptor (BCR) engagement. BLNK acts at the interface between BCR-associated Syk and downstream signaling cascades.

  1. Cheng, A.M. and Chan, A.C. (1997) Curr Opin Immunol 9, 528-33.
  2. Chu, D.H. et al. (1998) Immunol Rev 165, 167-80.
  3. Yamanashi, Y. et al. (1989) Proc Natl Acad Sci U S A 86, 6538-42.
  4. Khan, W.N. (2001) Immunol Res 23, 147-56.
  5. Tedder, T.F. et al. (1997) Immunity 6, 107-18.
  6. Kurosaki, T. (1997) Curr Opin Immunol 9, 309-18.
  7. Yamanashi, Y. et al. (1991) Science 251, 192-4.
  8. Lewis, C.M. et al. (2001) Curr Opin Immunol 13, 317-25.
  9. Buhl, A.M. and Cambier, J.C. (1999) J Immunol 162, 4438-46.
  10. Burkhardt, A.L. et al. (1991) Proc Natl Acad Sci U S A 88, 7410-4.
  11. Salim, K. et al. (1996) EMBO J 15, 6241-50.
  12. Fujimoto, M. et al. (2000) Immunity 13, 47-57.
  13. Turner, M. et al. (2000) Immunol Today 21, 148-54.
  14. Ren, C.L. et al. (1994) J Exp Med 179, 673-80.
  15. Rameh, L.E. et al. (1997) J Biol Chem 272, 22059-66.
  16. Coopman, P.J. et al. (2000) Nature 406, 742-7.
  17. Várnai, P. et al. (1999) J Biol Chem 274, 10983-9.
  18. Rawlings, D.J. et al. (1996) Science 271, 822-5.
  19. Park, H. et al. (1996) Immunity 4, 515-25.
  20. Zhang, J. et al. (2000) J Biol Chem 275, 35442-7.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
    XP is a registered trademark of Cell Signaling Technology, Inc.
    U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.
    All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

    Limited Uses

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    Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.