Cell Signaling(CST) 14471  PathScan® Intracellular Signaling Membrane Array Kit (Chemiluminescent Readout)

产品编号 14471 | Cell Signaling(CST)

• 产品描述




Kit Includes	Quantity	Applications	Reactivity	Homology†	MW (kDa)	Isotype
Array Membranes - Intracellular Signaling Array Kit (Membrane)	4 ea
Array 8-Well Dish - Membrane Array	1 ea
Array Diluent Buffer - Membrane Array	15 ml
Detection Ab Cocktail (10X) - Intracellular Signaling Array Kit (Membrane)	650 µl
HRP-Linked Streptavidin (10X) - Membrane Array	650 µl
SignalFire™ ECL Reagent #6883	6 ml
PathScan® Sandwich ELISA Lysis Buffer (1X) #7018	30 ml
20X Array Wash Buffer	15 ml
Chemiluminescent Development Folder	2




Specificity / Sensitivity

• PathScan^® Intracellular Signaling Membrane Array Kit (Chemiluminescent Readout) detects the indicated cellular proteins and signaling nodes only when phosphorylated or cleaved at the specified residues (see Figure 1). No significant cross-reactivity has been observed between targets. This kit is optimized for cell lysates diluted to a total protein concentration between 0.1 and 0.5 mg/ml (see kit protocol). This kit detects proteins from the indicated species as determined through in-house testing, but may also detect homologous proteins from other species.

Description

• The PathScan^® Intracellular Signaling Membrane Array Kit (Chemiluminescent Readout) is a nitrocellulose membrane antibody array founded upon the sandwich immunoassay principle. The array kit allows for the simultaneous detection of 18 important and well-characterized signaling molecules when phosphorylated or cleaved. Target-specific capture antibodies have been spotted in duplicate onto individual nitrocellulose membranes. Each kit contains 4 membranes, allowing the user to test up to 4 samples and generate 72 data points in a single experiment. Cell lysate is incubated on the membrane followed by a biotinylated detection antibody cocktail. Streptavidin-conjugated HRP and a chemiluminescent reagent are then used to visualize the bound detection antibody by chemiluminescence. An image of the membrane can be captured with either a digital imaging system or standard chemiluminescent film. The image can be analyzed visually or the spot intensities quantified using array analysis software.

Background

• HSP27 is a mediator of cell stress that confers resistance to adverse environmental change. HSP27 is phosphorylated at Ser78 within the p38 MAPK pathway.
• p53 plays an important role in cellular response to DNA damage and other genomic aberrations. Phosphorylation of p53 at Ser15 by ATM/ATR or DNA-PK in response to DNA damage leads to its stabilization and accumulation.
• Caspase-3 is a critical executor of apoptosis. Caspase-3 is activated by endoproteolytic cleavage at Asp175 and exerts its pro-apoptotic activity through cleavage of multiple cellular targets. PARP, an enzyme that is involved in DNA repair, is one of the main substrates of activated caspase-3. Cleavage at Asp214 leads to PARP inactivation. Increased levels of cleaved caspase-3 and cleaved PARP are reliable indicators of apoptosis.

Manning, B.D. and Cantley, L.C. (2007) Cell 129, 1261-74.

Huang, J. and Manning, B.D. (2009) Biochem Soc Trans 37, 217-22.

Dufner, A. and Thomas, G. (1999) Exp Cell Res 253, 100-9.

Hardie, D.G. et al. (2012) Nat Rev Mol Cell Biol 13, 251-62.

Rubinfeld, H. and Seger, R. (2005) Mol Biotechnol 31, 151-74.

Keshet, Y. and Seger, R. (2010) Methods Mol Biol 661, 3-38.

Cuadrado, A. and Nebreda, A.R. (2010) Biochem J 429, 403-17.

Brognard, J. and Hunter, T. (2011) Curr Opin Genet Dev 21, 4-11.

Hunter, T. (2009) Curr Opin Cell Biol 21, 140-6.

Manning, G. et al. (2002) Science 298, 1912-34.

Kurokawa, M. and Kornbluth, S. (2009) Cell 138, 838-54.

Shi, Y. (2004) Cell 117, 855-8.

Boatright, K.M. and Salvesen, G.S. (2003) Curr Opin Cell Biol 15, 725-31.

Application References

• Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Companion Products

12622 PathScan^® EGFR Signaling Antibody Array Kit (Chemiluminescent Readout)

12785 PathScan^® EGFR Signaling Antibody Array Kit (Fluorescent Readout)

12856 PathScan^® Stress and Apoptosis Signaling Antibody Array Kit (Chemiluminescent Readout)

12923 PathScan^® Stress and Apoptosis Signaling Antibody Array Kit (Fluorescent Readout)

13047 PathScan^® Th1/Th2/Th17 Cytokine Antibody Array Kit (Chemiluminescent Readout)

13124 PathScan^® Th1/Th2/Th17 Cytokine Antibody Array Kit (Fluorescent Readout)

7744 PathScan^® Intracellular Signaling Array Kit (Fluorescent Readout)

7949 PathScan^® RTK Signaling Antibody Array Kit (Fluorescent Readout)

7982 PathScan^® RTK Signaling Antibody Array Kit (Chemiluminescent Readout)

9474 PathScan^® Akt Signaling Antibody Array Kit (Chemiluminescent Readout)

9700 PathScan^® Akt Signaling Antibody Array Kit (Fluorescent Readout)

常见问题及解答

如何选择抗体

我看到很多抗体都加了一个XP®，XP® 是什么意思？

有些抗体名称里有个括号，括号里面是什么意思？

我怎么知道有没有我需要的蛋白？

实验疑难问题解答

WB封闭用牛奶好还是BSA好？二抗稀释液用什么？

做磷酸化抗体的时候一定要用脱脂奶粉稀释吗？

我的抗体的浓度是多少？

在组织样本中如何操作

我用的是组织样本，该如何破碎？

CST 的抗体是否适用于全组织样本裂解物？

抗体芯片

抗体芯片的保质期是多久？有多个孔，我一次用不了那么多，该如何保存？

细胞因子

人细胞因子适用于老鼠吗？

应该怎样配制PH3.0，20 mM的柠檬酸？

在我使用蛋白时是否应该使用载体蛋白？

siRNA

siRNA有两个或多个的情况，该如何选择？哪个效果好？

ELISA标准品

ELISA 有标准品吗？我如果买了其他公司的标准品，可不可以配套使用？

定制抗体

需要更进一步的帮助吗？

我能够订购一个定制配方的抗体吗？

CST能够定制我的抗体吗？

储存运输及发表文章

CST如何确保抗体的质量？

我该如何储存我的抗体？多高的温度会影响抗体的稳定性？

我需要发表文章，需要知道你们的产地，该如何写?

公司介绍

• CST公司是一家由研究型科学家创建于1999年的家族式私营企业，在全球范围内拥有超过400多名员工。CST活跃于应用系统生物学研究，尤其是与癌症相关的领域。作为一家始终致力于科学研究的公司，CST深知抗体的高特异性和批次间性能的一致性至关重要。因此我们坚持自己生产所有的抗体，并在多种应用中进行严格的验证。同时，正是生产这些抗体的CST科学家为客户提供技术支持，帮助他们设计实验，解决问题以获得最可靠的结果。我们之所以这样做，是因为如果我们做实验，我们也想要得到同样的服务。事实上，CST的确是这样的。
• Uante,您的科研物资战略合作伙伴！

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