Dropping with the temps: Cool deals on CST mAbs | Learn More >>
72159
B Cell Signaling Antibody Sampler Kit II
Primary Antibodies

B Cell Signaling Antibody Sampler Kit II #72159

IHC-P (paraffin) Image 1

Immunohistochemical analysis of paraffin-embedded human lymphoma using CD79A (D1X5C) XP® Rabbit mAb.

Learn more about how we get our images
Western Blotting Image 2

Western blot analysis of extracts from various cell lines using CD79A (D1X5C) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).

Learn more about how we get our images
Western Blotting Image 3

Western blot analysis of extracts from Ramos cells untreated or treated with anti-human IgM (12 µg/ml for 10 minutes), using Phospho-CD79A (Tyr182) Antibody (upper) or CD79A Antibody #3351 (lower).

Learn more about how we get our images
Western Blotting Image 4

Western blot analysis of extracts from various cell lines using CD19 (D4V4B) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower). As expected, CD19 protein is not detected in either U-937 cells or Raw 264.7 cells.

Learn more about how we get our images
IHC-Leica® Bond™ Image 5

Immunohistochemical analysis of paraffin-embedded human endometrioid carcinoma using CD19 (D4V4B) XP® Rabbit mAb performed on the Leica® Bond™ Rx.

Learn more about how we get our images
Western Blotting Image 6

Western blot analysis of SDS extracts from untreated or anti-human IgM treated (12 µg/ml for 2 minutes) Ramos cells using Phospho-CD19 (Tyr531) Antibody (upper) and control CD19 Antibody #3574 (lower).

Learn more about how we get our images
Western Blotting Image 7

Western blot analysis of extracts from Daudi cells, serum-starved overnight, then vehicle-treated (lane 1), treated with anti-human IgM (12 μg/ml, 10 min; lane 2), or pre-treated with Ibrutinib #16483 (1 μM, 60 min) prior to anti-IgM treatment (lane 3), using Phospho-Btk (Tyr23) (D1D2Z) Rabbit mAb (upper) or Btk (D6H5) Rabbit mAb #8547 (lower).

Learn more about how we get our images
Western Blotting Image 8

Western blot analysis of extracts from various cell lines using Syk (D3Z1E) XP® Rabbit mAb.

Learn more about how we get our images
Western Blotting Image 9

Western blot analysis of extracts from Ramos cells, untreated or treated with anti-IgM, using Phospho-Syk (Tyr525/526) (C87C1) Rabbit mAb (upper) or Syk Antibody #2712 (lower).

Learn more about how we get our images
Western Blotting Image 10

Western blot analysis of SDS extracts from control or anti-human IgM treated (12 µg/ml for 2 minutes) Ramos cells using Phospho-BLNK (Tyr96) Antibody.

Learn more about how we get our images
Western Blotting Image 11

Western blot analysis of extracts from various cell lines using Btk (D3H5) Rabbit mAb.

Learn more about how we get our images
Flow Cytometry Image 12

Human whole blood was fixed, lysed, and permeabilized as per the Cell Signaling Technology Flow Alternate Protocol and stained using Btk (D3H5) Rabbit mAb. Samples were co-stained using CD3-PE and CD19-APC to distinguish T and B cell subpopulations, respectively. B (red) and T (blue) cell population gates were applied to a histogram depicting the mean fluorescence intensity of Btk. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

Learn more about how we get our images
IHC-Leica® Bond™ Image 13

Immunohistochemical analysis of paraffin-embedded human squamous cell lung carcinoma using Btk (D3H5) Rabbit mAb performed on the Leica® Bond™ Rx.

Learn more about how we get our images
Western Blotting Image 14

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

Learn more about how we get our images
Western Blotting Image 15

Western blot analysis of extracts from Ramos cells, untreated (-) or PNGase F-treated (+), using CD79A (D1X5C) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).

Learn more about how we get our images
Flow Cytometry Image 16

Flow cytometric analysis of Ramos cells, untreated (blue) or IgM-treated (green), using Phospho-CD79A (Tyr182) Antibody.

Learn more about how we get our images
Western Blotting Image 17

Western blot analysis of extracts from Daudi cells, untreated (-) or treated with PNGase F (+), using CD19 (D4V4B) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).

Learn more about how we get our images
IP Image 18

Immunoprecipitation of Phospho-Btk (Tyr223) from Ramos cells, serum-starved followed by treatment with anti-human IgM (12 μg/ml, 10 min) using Phospho-Btk (Tyr223) (D1D2Z) Rabbit mAb. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotyope Control #3900, lane 3 is Phospho-Btk (Tyr223) (D1D2Z) Rabbit mAb (1:50), and lane 4 is Phospho-Btk (Tyr223) (D1D2Z) Rabbit mAb (1:100). Western blot analysis was performed using Phospho-Btk (Tyr223) (D1D2Z) Rabbit mAb. Anti-rabbit IgG, HRP-linked Antibody #7074 was used as the secondary antibody.

Learn more about how we get our images
IP Image 19

Immunoprecipitation of Syk protein from SR cell extracts, using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or Syk(D3Z1E) XP® Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using Syk (D3Z1E) XP® Rabbit mAb

Learn more about how we get our images
Flow Cytometry Image 20

Flow cytometric analysis of Ramos cells, untreated (blue) or treated with anti-IgM (green), using Phospho-Syk(Tyr525/526) (C87C1) Rabbit mAb. Anti-rabbit IgG (H+L), F(ab')2 Fragment (PE Conjugate) #8885 was used as a secondary antibody.

Learn more about how we get our images
IHC-P (paraffin) Image 21

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using Btk (D3H5) Rabbit mAb. Note staining of inflammatory cells.

Learn more about how we get our images
IHC-P (paraffin) Image 22

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using CD79A (D1X5C) XP® Rabbit mAb.

Learn more about how we get our images
IF-IC Image 23

Confocal immunofluorescent analysis of Ramos cells, untreated (left) or IgM-treated (middle), and Jurkat cells (right) using Phospho-CD79A (Tyr182) Antibody (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Learn more about how we get our images
IHC-P (paraffin) Image 24

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using CD19 (D4V4B) XP® Rabbit mAb.

Learn more about how we get our images
IHC-P (paraffin) Image 25

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Syk (D3Z1E) XP® Rabbit mAb.

Learn more about how we get our images
IF-IC Image 26

Confocal immunofluorescent analysis of Ramos cells, serum-starved (overnight; left) or IgM-treated (12 ug/ml, 2 minutes; right), using Phospho-Syk (Tyr525/526) (C87C1) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Learn more about how we get our images
IHC-P (paraffin) Image 27

Immunohistochemical analysis of paraffin-embedded human B-cell lymphoma using Btk (D3H5) Rabbit mAb.

Learn more about how we get our images
IHC-P (paraffin) Image 28

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using CD19 (D4V4B) XP® Rabbit mAb.

Learn more about how we get our images
IHC-P (paraffin) Image 29

Immunohistochemical analysis of paraffin-embedded human lymph node using Syk (D3Z1E) XP® Rabbit mAb.

Learn more about how we get our images
IHC-P (paraffin) Image 30

Immunohistochemical analysis of paraffin-embedded mouse colon using Btk (D3H5) Rabbit mAb.

Learn more about how we get our images
IHC-P (paraffin) Image 31

Immunohistochemical analysis of paraffin-embedded human lung carcinoma using CD79A (D1X5C) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).

Learn more about how we get our images
IHC-P (paraffin) Image 32

Immunohistochemical analysis of paraffin-embedded human lymphoma using CD19 (D4V4B) XP® Rabbit mAb.

Learn more about how we get our images
IHC-P (paraffin) Image 33

Immunohistochemical analysis of paraffin-embedded mouse spleen using Syk (D3Z1E) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).

Learn more about how we get our images
IHC-P (paraffin) Image 34

Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma using Btk (D3H5) Rabbit mAb. Note staining of inflammatory cells.

Learn more about how we get our images
Flow Cytometry Image 35

Flow cytometric analysis of human whole blood lymphocytes using CD79A (D1X5C) XP® Rabbit mAb. Cells were co-stained with CD19-APC and CD3-PE antibodies to allow for CD79A analysis in B cells (green) and T cells (blue), respectively. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

Learn more about how we get our images
IHC-P (paraffin) Image 36

Immunohistochemical analysis of paraffin-embedded mouse spleen using CD19 (D4V4B) XP® Rabbit mAb.

Learn more about how we get our images
Flow Cytometry Image 37

Flow cytometric analysis of RL-7 cells using Syk (D3Z1E) XP® Rabbit mAb (blue) compared to Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (red). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 647 Conjugate) #4414 was used as a secondary antibody.

Learn more about how we get our images
IHC-P (paraffin) Image 38

Immunohistochemical analysis of paraffin-embedded cell pellets, Ramos(left) or Jurkat (right), using Btk (D3H5) Rabbit mAb.

Learn more about how we get our images
IHC-P (paraffin) Image 39

Immunohistochemical analysis of paraffin-embedded human tonsil using CD19 (D4V4B) XP® Rabbit mAb.

Learn more about how we get our images
Product Includes Quantity Applications Reactivity MW(kDa) Isotype
CD79A (D1X5C) XP® Rabbit mAb 13333 20 µl
  • WB
  • IHC
  • F
H 45-55 Rabbit IgG
Phospho-CD79A (Tyr182) Antibody 5173 20 µl
  • WB
  • IP
  • IF
  • F
H 45-55 Rabbit 
CD19 (D4V4B) XP® Rabbit mAb 90176 20 µl
  • WB
  • IP
  • IHC
H M 95 Rabbit IgG
Phospho-CD19 (Tyr531) Antibody 3571 20 µl
  • WB
  • IP
H 95 Rabbit 
Phospho-Btk (Tyr223) (D1D2Z) Rabbit mAb 87457 20 µl
  • WB
  • IP
H M 78 Rabbit IgG
Syk (D3Z1E) XP® Rabbit mAb 13198 20 µl
  • WB
  • IP
  • IHC
  • F
H M R 72 Rabbit IgG
Phospho-Syk (Tyr525/526) (C87C1) Rabbit mAb 2710 20 µl
  • WB
  • IP
  • IF
  • F
H 72 Rabbit IgG
Phospho-BLNK (Tyr96) Antibody 3601 20 µl
  • WB
  • IP
H 68, 70 Rabbit 
Btk (D3H5) Rabbit mAb 8547 20 µl
  • WB
  • IP
  • IHC
  • F
H M 77 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

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.

All antibodies contained in this kit detect endogenous levels of their respective target protein.

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide and are purified by protein A and peptide affinity chromatography. Monoclonal antibodies are produced by immunizing animals with recombinant human proteins or synthetic peptides.

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.
Entrez-Gene Id
29760 , 695 , 930 , 973 , 6850
Swiss-Prot Acc.
Q8WV28 , Q06187 , P15391 , P11912 , P43405
For Research Use Only. Not For Use In Diagnostic Procedures.

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.

Upstream / Downstream

pathwayImage

Explore pathways related to this product.