Revision 6

#13198Store at -20C

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.
Applications:

WB, W-S, IP, IHC-P, IF-IC, FC-FP

REACTIVITY:

H M R

SENSITIVITY:

Endogenous

MW (kDa):

72

Source/Isotype:

Rabbit IgG

UniProt ID:

#P43405

Entrez-Gene Id:

6850

Product Information

Product Usage Information

Application Dilution
Western Blotting 1:1000
Simple Western™ 1:10 - 1:50
Immunoprecipitation 1:50
Immunohistochemistry (Paraffin) 1:300
Immunofluorescence (Immunocytochemistry) 1:800 - 1:1600
Flow Cytometry (Fixed/Permeabilized) 1:100

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.

For a carrier free (BSA and azide free) version of this product see product #27543.

Specificity / Sensitivity

Syk (D3Z1E) XP® Rabbit mAb recognizes endogenous levels of total Syk protein.

Species Reactivity:

Human, Mouse, Rat

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues around Asn463 of human Syk protein.

Background

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 of a role for Syk in nonimmune cells and investigators have indicated that Syk is a potential tumor suppressor in human breast carcinomas (5). Tyr323 is a negative regulatory phosphorylation site within the SH2-kinase linker region in Syk. Phosphorylation at Tyr323 provides a direct binding site for the TKB domain of Cbl (6,7). Tyr352 of Syk is involved in the association of PLCγ1 (8). Tyr525 and Tyr526 are located in the activation loop of the Syk kinase domain; phosphorylation at Tyr525/526 of human Syk (equivalent to Tyr519/520 of mouse Syk) is essential for Syk function (9).

  1. Cheng, A.M. and Chan, A.C. (1997) Curr Opin Immunol 9, 528-33.
  2. Kurosaki, T. (1997) Curr Opin Immunol 9, 309-18.
  3. Chu, D.H. et al. (1998) Immunol Rev 165, 167-80.
  4. Turner, M. et al. (2000) Immunol Today 21, 148-54.
  5. Coopman, P.J. et al. (2000) Nature 406, 742-7.
  6. Deckert, M. et al. (1998) J Biol Chem 273, 8867-74.
  7. Rao, N. et al. (2001) EMBO J 20, 7085-95.
  8. Law, C.L. et al. (1996) Mol Cell Biol 16, 1305-15.
  9. Zhang, J. et al. (2000) J Biol Chem 275, 35442-7.

Species Reactivity

Species reactivity is determined by testing in at least one approved application (e.g., western blot).

Western Blot Buffer

IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.

Applications Key

WB: Western Blotting W-S: Simple Western™ IP: Immunoprecipitation IHC-P: Immunohistochemistry (Paraffin) IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized)

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
Alexa Fluor is a registered trademark of Life Technologies Corporation.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

限制使用

除非 CST 的合法授书代表以书面形式书行明确同意,否书以下条款适用于 CST、其关书方或分书商提供的书品。 任何书充本条款或与本条款不同的客书条款和条件,除非书 CST 的合法授书代表以书面形式书独接受, 否书均被拒书,并且无效。

专品专有“专供研究使用”的专专或专似的专专声明, 且未专得美国食品和专品管理局或其他外国或国内专管机专专专任何用途的批准、准专或专可。客专不得将任何专品用于任何专断或治专目的, 或以任何不符合专专声明的方式使用专品。CST 专售或专可的专品提供专作专最专用专的客专,且专用于研专用途。将专品用于专断、专防或治专目的, 或专专售(专独或作专专成)或其他商专目的而专专专品,均需要 CST 的专独专可。客专:(a) 不得专独或与其他材料专合向任何第三方出售、专可、 出借、捐专或以其他方式专专或提供任何专品,或使用专品制造任何商专专品,(b) 不得复制、修改、逆向工程、反专专、 反专专专品或以其他方式专专专专专品的基专专专或技专,或使用专品开专任何与 CST 的专品或服专专争的专品或服专, (c) 不得更改或专除专品上的任何商专、商品名称、徽专、专利或版专声明或专专,(d) 只能根据 CST 的专品专售条款和任何适用文档使用专品, (e) 专遵守客专与专品一起使用的任何第三方专品或服专的任何专可、服专条款或专似专专

Revision 6
#13198

Syk (D3Z1E) XP® Rabbit mAb

Western Blotting Image 1: Syk (D3Z1E) XP® Rabbit mAb Expand Image
使用 Syk (D3Z1E) XP® Rabbit mAb 对不同细胞系的提取物进行蛋白质印迹分析。
Western Blotting Image 2: Syk (D3Z1E) XP® Rabbit mAb Expand Image
使用 Syk (D3Z1E) XP® Rabbit mAb(上图)对 THP-1 WT(左图)或 SYK KO(右图)细胞的提取物进行蛋白质印迹法分析。用 Ponceau S 染色的膜进行总蛋白标准化(下图)。这些数据由 YCharOS Inc. 提供,该公司是一家开放科学公司,作为 CST 科学家生成的验证数据的合作伙伴,其使命是表征市售抗体。
Western Blotting Image 1: Syk (D3Z1E) XP® Rabbit mAb Expand Image
使用 Syk (D3Z1E) XP® Rabbit mAb #13198 对 Raji 细胞的裂解物 (0.1 mg/mL) 进行 Simple Western™ 分析。虚拟泳道式图像(左图)显示一抗稀释比例为 1:10 和 1:50 时的靶标条带(如图所示)。对应的电泳图(右图)为一抗稀释比例在 1:10(蓝线)和 1:50(绿线)时沿毛细血管内分子量的化学发光结果。在还原条件下,使用 12-230 kDa 分离模块在 ProteinSimple(BioTechne 品牌)的 Jess™ Simple Western 仪器上进行该实验。
Immunoprecipitation Image 1: Syk (D3Z1E) XP® Rabbit mAb Expand Image
使用 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(泳道 2)或 Syk(D3Z1E) XP® Rabbit mAb(泳道 3)对 SR 细胞提取物 Syk 蛋白进行免疫沉淀。泳道 1 是 10% 输入对照。使用 Syk (D3Z1E) XP® Rabbit mAb 进行蛋白质印迹分析。
Immunohistochemistry Image 1: Syk (D3Z1E) XP® Rabbit mAb Expand Image
使用 Syk (D3Z1E) XP® Rabbit mAb 对石蜡包埋的人乳腺癌细胞进行免疫组织化学分析。
Immunohistochemistry Image 2: Syk (D3Z1E) XP® Rabbit mAb Expand Image
使用 Syk (D3Z1E) XP® Rabbit mAb 对石蜡包埋的人淋巴结细胞进行免疫组织化学分析。
Immunohistochemistry Image 3: Syk (D3Z1E) XP® Rabbit mAb Expand Image
在有对照肽(左图)或抗原特异性肽(右图)的情况下,使用 Syk (D3Z1E) XP® Rabbit mAb 对石蜡包埋的小鼠脾脏细胞进行免疫组织化学分析。
Immunofluorescence Image 1: Syk (D3Z1E) XP® Rabbit mAb Expand Image
使用 Syk (D3Z1E) XP® Rabbit mAb(绿色)、DyLight 650 Phalloidin #12956(红色)和 DAPI #4083(蓝色),对 SW620 细胞(左图,阳性)和 ACHN 细胞(右图,阴性)进行共聚焦免疫荧光分析。
Flow Cytometry Image 1: Syk (D3Z1E) XP® Rabbit mAb Expand Image
以 Rabbit (DA1E) mAb IgG XP® Isotype Control #3900(红色)作为对照,使用 Syk (D3Z1E) XP® Rabbit mAb(蓝色)对 RL 细胞进行流式细胞分析。Anti-rabbit IgG (H+L)、F(ab')2 Fragment (Alexa Fluor® 647 Conjugate) #4414 作为二抗。