Cat. # | Size | Price | Inventory |
---|---|---|---|
55147S | 100 µl |
REACTIVITY | H |
SENSITIVITY | Endogenous |
MW (kDa) | 45-53 |
Source/Isotype | Rabbit IgG |
Product Information
Application | Dilution |
---|---|
Western Blotting | 1:1000 |
Immunofluorescence (Immunocytochemistry) | 1:100 |
Flow Cytometry (Fixed/Permeabilized) | 1:200 - 1:800 |
Flow Cytometry (Live) | 1:50 - 1:200 |
For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody product webpage for recommended antibody dilution.
From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: #12957 Western Blotting Application Solutions Kit
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Loading of prestained molecular weight markers (#59329, 10 µl/lane) to verify electrotransfer and biotinylated protein ladder (#7727, 10 µl/lane) to determine molecular weights are recommended.
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised June 2020
Protocol Id: 10
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalently purified water.
Recommended Fluorochrome-conjugated Anti-Rabbit secondary antibodies:
NOTE: Cells should be grown, treated, fixed and stained directly in multi-well plates, chamber slides or on coverslips.
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 December 2010
Protocol Id: 3
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:404
注:使用反渗透去离子水 (RODI) 或同等级别的水配制溶液。
注:在您的实验中加入荧光细胞染料(包括活力指示染料、DNA 染料等)时,请参考染料产品网页,了解建议的实验步骤。访问 www.cellsignal.com,了解经验证用于流式细胞术的细胞染料完整列表。
注:使用血细胞计数器或备选方法计数细胞。
注:如果使用全血,则需裂解红血细胞,并在免疫染色之前通过离心分离洗涤。
注:人 Fc 受体与兔 IgG 发生交叉反应。当感兴趣的细胞表达高水平的 Fc 受体蛋白(例如,巨噬细胞/单核细胞谱系)时,在用兔抗体进行免疫染色之前,用人 Fc 块预孵育活细胞。
注:最佳离心条件会根据细胞类型和试剂容量变动。一般,1-5 分钟 150-300g 将足以使细胞沉淀下来。
修订时间 2022 年 1 月
实验步骤编号:1865
人
使用与人组织因子/CD142 蛋白的 Glu123 周围的残基相对应的合成肽对动物进行免疫接种来产生单克隆抗体。
组织因子 (TF)/CD142 (凝血因子III /促凝血酶原激酶) 是一种 I 型跨膜糖蛋白,作为血液凝固因子 VII 和 VIIa 的细胞表面受体和辅助因子而发挥作用,因此在止血和血栓形成中具有核心作用 (1)。TF:VIIa 受体-配体复合体被广泛地认为是外在血凝蛋白激酶级联反应的启动因子,它最终可导致纤维蛋白和凝血酶生成 (1)。作为 II 型细胞因子受体超家族成员,TF 也被证明可以参与结合 VIIa 因子后的 PI3K (2) 与 MAPK (3) 信号转导级联反应,以驱动细胞迁移、生长和增殖等细胞反应。尽管在生理条件下,TF 的作用是响应组织损伤时调血液凝固,但 TF 在病理状况(比如肿瘤发生)下也有意义。事实上,TF 在结直肠癌、乳腺癌、胰腺癌和多形性胶质母细胞瘤中均异常表达 (4)。它已被证明可以促进肿瘤血管生成、肿瘤生长、转移和静脉血栓形成 (5)。鉴于 TF 过表达与许多类型的实体肿瘤有关,作为一个潜在的治疗靶标,它引起了大量关注。
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。