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THIS PRODUCT IS DISCONTINUED
Fast Activated Cell-based ELISA (FACE™) Kits provide a simple, sensitive method for detecting protein phosphorylation directly in the cell, without making extracts or performing electrophoresis and membrane blotting. These 96-well, high-throughput assays are available in both colorimetric and chemiluminescent formats for over 20 different targets (see list at right). For complete details, click the FACE™ Method tab below.
FACE JNK Kits provide 96 rxns each of 2 antibodies that enable you to monitor and compare the levels of both phosphorylated and total JNK (c-Jun N-terminal Kinase). The phospho-JNK antibody recognizes JNK1 only when phosphorylated at Thr183/Tyr185. The total-JNK antibody recognizes JNK1, JNK2 and JNK3 regardless of their phosphorylation states. Click the JNK Info tab below for data and more information.
| Name | Format | Cat No. | 価格 (税抜) | |
|---|---|---|---|---|
| FACE™ JNK | 1 x 96 rxns | 48110 | Discontinued | |
| 5 x 96 rxns | 48610 | Discontinued | ||
| FACE™ JNK Chemi | 1 x 96 rxns | 48210 | Discontinued | |
| 5 x 96 rxns | 48710 | Discontinued | ||
| FACE™ JNK Manual |
| FACE™ Profile |
| Cell Biology Products Brochure |
| IsoCyte™ Application Note – Phospho-Protein Detection |
| MSDS: Sodium Azide |
| MSDS: Sulphuric Acid |
| MSDS: Thimersol |
Figure 1: Colorimetric measurement of phosphorylated and total JNK.Murine macrophage 4/4 cells were cultured in 96-well plates and serum-starved for 16 hours before stimulation. Cells were stimulated at the indicated concentrations of anisomycin for 15 minutes and then fixed. Total and phospho-JNK were each assayed in triplicate using the phospho and total JNK antibodies from the FACE JNK Kit. Data was plotted after correction for cell number (performed through use of Crystal Violet). Note that the induction treatment did not affect the overall level of total JNK.
Antibody Specificities
The phospho-JNK antibody is specific for phosphorylated JNK and was raised against a phospho-peptide that corresponds to the sequence surrounding Thr183/Tyr185 of human JNK. It recognizes JNK1 only when phosphorylated at Thr183/Tyr185, and JNK2 and JNK3 only when phosphorylated at equivalent sites. The total-JNK antibody recognizes JNK1, JNK2 and JNK3 regardless of the phosphorylation state.
JNK Overview
JNK (c-Jun N-terminal kinase), also called stress activated protein kinase (SAPK), is a member of the serine/threonine MAP kinase family. MAP kinase signaling cascades are critically important in translating the signals received at the plasma membrane into changes in cellular physiology and gene expression. JNK is activated in response to a variety of stimuli, including inflammatory cytokines, growth factors and cellular stresses such as UV-light. Because of JNK’s role in basic cellular processes such as inflammation and apoptosis, an understanding of the JNK signaling cascade should speed the development of therapeutics.
FACE™ の使用法
96ウェルプレートで細胞を培養し, 目的の経路を誘導する刺激を与えます。刺激を与えた後, 細胞を固定することでリン酸化を含むタンパク質修飾が保持されます。ブロッキング反応の後, それぞれのウェルに目的タンパク質に特異的な一次抗体, 続いてHRP標識二次抗体を加えます。その後, 発色試薬を加えることにより, 比色定量または化学発光による定量を簡便に行うことができます (図1)。また, キット付属のCrystal Violetを用いて細胞数を補正することができます。それぞれのFACE Kitには目的タンパク質全体とリン酸化型を認識する一次抗体が含まれます。これにより, 細胞数と細胞中の目的タンパク質の総量に対してのリン酸化型タンパク質量を解析することができます。FACEはリン酸化型タンパク質を解析することのできるキットで, 定量的かつ再現性のある結果が得られます。
図1: FACE の操作法
Contents & Storage
Two (or ten) 96-well plates for culturing cells, 96 (or 5 x 96) rxns each of two primary antibodies (1 phospho-specific, 1 specific for native protein), HRP-conjugated secondary antibody, Quenching Solution, 1X Antibody Blocking Buffer, 1X Antibody Dilution Buffer, 10X PBS, 10% Triton X-100, 1% SDS Solution, Developing and Stop Solutions, and Crystal Violet Cell Quantification Solution. Storage conditions vary from room temperature to -20°C, see manual for details. All reagents are guaranteed stable for 6 months when stored properly.
