Cat. # | Size | Price | Inventory |
---|---|---|---|
7497S | 100 µl (50 tests) |
REACTIVITY | H M R Mk |
SENSITIVITY | Endogenous |
MW (kDa) | |
Source/Isotype | Rabbit IgG |
Product Information
Application | Dilution |
---|---|
Immunofluorescence (Immunocytochemistry) | 1:100 |
Flow Cytometry (Fixed/Permeabilized) | 1:50 |
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
NOTE: Cells should be grown, treated, fixed and stained directly in multi-well plates, chamber slides or on coverslips.
Aspirate liquid, then cover cells to a depth of 2–3 mm with 4% formaldehyde diluted in 1X PBS.
NOTE: Formaldehyde is toxic, use only in a fume hood.
NOTE: All subsequent incubations should be carried out at room temperature unless otherwise noted in a humid light-tight box or covered dish/plate to prevent drying and fluorochrome fading.
posted November 2006
revised November 2013
Protocol Id: 182
All reagents required for this protocol may be efficiently purchased together in our Intracellular Flow Cytometry Kit (Methanol) #13593, or individually using the catalog numbers listed below.
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
NOTE: When including fluorescent cellular dyes in your experiment (including viability dyes, DNA dyes, etc.), please refer to the dye product page for the recommended protocol. Visit www.cellsignal.com for a full listing of cellular dyes validated for use in flow cytometry.
NOTE: Adherent cells or tissue should be dissociated and in single-cell suspension prior to fixation.
NOTE: Optimal centrifugation conditions will vary depending upon cell type and reagent volume. Generally, 150-300g for 1-5 minutes will be sufficient to pellet the cells.
NOTE: If using whole blood, lyse red blood cells and wash by centrifugation prior to fixation.
NOTE: Antibodies targeting CD markers or other extracellular proteins may be added prior to fixation if the epitope is disrupted by formaldehyde and/or methanol. The antibodies will remain bound to the target of interest during the fixation and permeabilization process. However, note that some fluorophores (including PE and APC) are damaged by methanol and thus should not be added prior to permeabilization. Conduct a small-scale experiment if you are unsure.
NOTE: Count cells using a hemocytometer or alternative method.
posted July 2009
revised June 2020
实验步骤编号:407
人, 小鼠, 大鼠, 猴
使用与人 NDRG1 蛋白中 Thr346 周围残基相对应的合成磷酸肽,对动物进行免疫接种来产生单克隆抗体。
N-myc 下游调节基因 1 (NDRG1),也称为 Cap43、Drg1、RTP/rit42、Proxy-1,是 NDRG 家族成员之一,由在生长、分化和细胞存活中起作用的四个成员 (NDRG1-4) 组成 (1-5)。NDRG1 广泛表达,高度响应于各种应激信号,包括 DNA 损伤 (4)、低氧 (5) 和镍与钙水平升高 (2)。NDRG1 表达在 N-myc 缺陷小鼠中升高,并受 N- 和 c-myc 负向调控 (1,6)。DNA 损伤期间,NDRG1 以 p53 依赖性方式诱导,并且对 p53 介导的凋亡必不可少 (4,7)。研究表明,NDRG1 在癌症进展中起到促进分化、抑制生长以及调节转移和血管生成的作用 (3,4,6,8,9)。已经证明 NDRG1 基因的无义突变导致遗传性运动和神经病变 (HMSNL),这获得众多展示 NDRG1 起到维护髓鞘和轴突存活作用的研究支持 (10,11)。NDRG1 在肥大细胞成熟过程中上调,其缺失导致过敏反应减弱(12)。NDRG1 和 NDRG2 都是 SGK1 的底物,不过未知 SGK1 介导的磷酸化的确切生理作用 (13)。NDRG1 在位点 Thr328、Ser330、Thr346、Thr356 和 Thr366 为 SGK1 磷酸化。SGK1 所致的磷酸化为 GSK-3 磷酸化 NDRG1 做好准备。
除非如以 CST 合法授权代表签署的书面形式另行明确同意,否则以下条款适用于 CST、其附属公司或其分销商提供的产品。除非 CST 合法授权代表以书面形式单独接受,否则任何附加于或异于此处所载条款和条件的客户条款和条件均被拒绝且无效。
产品用“仅供研究使用”或类似标示声明标示,并且尚未经 FDA 或其他国外或国内监管实体出于任何目的批准、准许或许可。客户不得出于任何诊断或治疗目的或以任何与产品标示声明相冲突的方式使用任何产品。CST 销售或许可的产品提供给作为最终用户的客户,且仅用于研究和开发用途。出于诊断、预防或治疗目的任何产品使用或出于转售(单独或作为成分)或其他商业目的的任何产品购买都要求来自 CST 的单独许可。客户 (a) 不得向任何第三方出售、许可、出借、捐赠或另行转让或提供任何本公司产品,无论单独或联合其他材料方式,或使用本公司产品制造任何商业产品,(b) 不得复制、修改、逆向工程、反编译、反汇编或另行尝试发现本公司产品的底层结构或技术,或出于开发与 CST 产品或服务竞争的任何产品或服务的目的使用本公司产品,(c) 不得从本公司产品改变或移除任何商标、商品名称、徽标、专利或版权声明或标记,(d) 仅应根据 CST 产品销售条款和任何适用文档使用本公司产品,以及 (e) 应就客户联系本公司产品所用的任何第三方产品或服务而言遵守任何许可、服务条款或类似协议。