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#72159 B Cell Signaling Antibody Sampler Kit II

 
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2019年1月17日11時55分 現在
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#72159T1 Kit115,000
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キット内容 容量 用途 種交差性 ホモロジー† 検出分子量 アイソタイプ
CD79A (D1X5C) XP® Rabbit mAb #13333 20 µl WB, IHC-P, F H 45-55 Rabbit IgG
Phospho-CD79A (Tyr182) Antibody #5173 20 µl WB, IP, IF-IC, F H M, R 45-55 Rabbit
CD19 (D4V4B) XP® Rabbit mAb #90176 20 µl WB, IP, IHC-Bond, IHC-P H, M B 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-P, F H, M, R 72 Rabbit IgG
Phospho-Syk (Tyr525/526) (C87C1) Rabbit mAb #2710 20 µl WB, IP, IF-IC, F H M, R 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-Bond, IHC-P, F H, M R, Hm, B, Dg, Pg, Hr 77 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl WB Goat

†Species predicted to react based on 100% sequence homology.

Applications Key: W=Western Blotting, IHC-P=Immunohistochemistry (Paraffin), F=Flow Cytometry, IP=Immunoprecipitation, IF-IC=Immunofluorescence (Immunocytochemistry), IHC-Bond=IHC-Leica® Bond™
Reactivity Key: H=Human, M=Mouse, R=Rat

特異性・感度
All antibodies contained in this kit detect endogenous levels of their respective target protein.
使用抗原
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.

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貯法
-20℃
社内データ

Western Blotting

Western Blotting

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.

Western Blotting

Western Blotting

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.

Western Blotting

Western Blotting

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).


Western Blotting

Western Blotting

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).

Western Blotting

Western Blotting

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).

Western Blotting

Western Blotting

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


Flow Cytometry

Flow Cytometry

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.

Western Blotting

Western Blotting

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

Western Blotting

Western Blotting

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


IHC-P (paraffin)

IHC-P (paraffin)

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

Western Blotting

Western Blotting

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.

IHC-Leica® Bond™

IHC-Leica® Bond™

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


IHC-Leica® Bond™

IHC-Leica® Bond™

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

Western Blotting

Western Blotting

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).

Flow Cytometry

Flow Cytometry

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


IHC-P (paraffin)

IHC-P (paraffin)

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

Flow Cytometry

Flow Cytometry

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.

IP

IP

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


Western Blotting

Western Blotting

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).

Western Blotting

Western Blotting

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).

IP

IP

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.


IF-IC

IF-IC

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).

IHC-P (paraffin)

IHC-P (paraffin)

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

IF-IC

IF-IC

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).


IHC-P (paraffin)

IHC-P (paraffin)

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

IHC-P (paraffin)

IHC-P (paraffin)

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

IHC-P (paraffin)

IHC-P (paraffin)

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


IHC-P (paraffin)

IHC-P (paraffin)

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

IHC-P (paraffin)

IHC-P (paraffin)

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

IHC-P (paraffin)

IHC-P (paraffin)

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


IHC-P (paraffin)

IHC-P (paraffin)

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

IHC-P (paraffin)

IHC-P (paraffin)

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).

IHC-P (paraffin)

IHC-P (paraffin)

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).


IHC-P (paraffin)

IHC-P (paraffin)

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

IHC-P (paraffin)

IHC-P (paraffin)

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

Flow Cytometry

Flow Cytometry

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.


Flow Cytometry

Flow Cytometry

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.

IHC-P (paraffin)

IHC-P (paraffin)

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

IHC-P (paraffin)

IHC-P (paraffin)

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


バックグラウンド

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.

使用例
製品をご使用いただいて研究を発表されましたら、ぜひお知らせください
 
関連製品
12630   SignalFire™ Plus ECL Reagent
12757   SignalFire™ Elite ECL Reagent
13198   Syk (D3Z1E) XP® Rabbit mAb
13333   CD79A (D1X5C) XP® Rabbit mAb
13953   Prestained Protein Marker, Broad Range (11-190 kDa)
2710   Phospho-Syk (Tyr525/526) (C87C1) Rabbit mAb
3571   Phospho-CD19 (Tyr531) Antibody
3601   Phospho-BLNK (Tyr96) Antibody
5173   Phospho-CD79A (Tyr182) Antibody
6883   SignalFire™ ECL Reagent
7003   20X LumiGLO® Reagent and 20X Peroxide
7074   Anti-rabbit IgG, HRP-linked Antibody
7075   Anti-biotin, HRP-linked Antibody
7727   Biotinylated Protein Ladder Detection Pack
8547   Btk (D3H5) Rabbit mAb
87457   Phospho-Btk (Tyr223) (D1D2Z) Rabbit mAb
90176   CD19 (D4V4B) XP® Rabbit mAb
9997   Tris Buffered Saline with Tween® 20 (TBST-10X)
9999   Nonfat Dry Milk

BOND is a trademark of Leica Biosystems Melbourne Pty. Ltd. No affiliation or sponsorship between CST and Leica Microsystems IR GmbH or Leica Biosystems Melbourne Pty. Ltd is implied.
Tween is a registered trademark of ICI Americas, Inc.
DRAQ5 is a registered trademark of Biostatus Limited.
SignalStain is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.
LEICA is a registered trade​mark of Leica Microsystems IR GmbH.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
LumiGLO is a registered trademark of Kirkegaard & Perry Laboratories.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.

本製品は試験研究用です。

B Cell Signaling Antibody Sampler Kit II

Immune Cell Signaling Pathways

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