Cat. # | Size | Price | Inventory |
---|---|---|---|
14047S | 100 µl (50 tests) |
REACTIVITY | H M R Mk |
SENSITIVITY | Endogenous |
MW (kDa) | |
Source/Isotype | Rabbit IgG |
Product Information
Application | Dilution |
---|---|
Immunofluorescence (Immunocytochemistry) | 1:800 |
Flow Cytometry (Fixed/Permeabilized) | 1:50 |
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalently purified water.
NOTE: Cells should be grown, treated, fixed and stained directly in multiwell 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 November 2006
revised December 2010
Protocol Id: 220
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
牛 , 猪
使用与人 Stat3 蛋白中 Gly700 周围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。
Stat3 转录因子是众多细胞因子和生长因子受体的重要信号转导 (1),对胎鼠发育必不可少 (2)。研究显示 Stat3 在多种人类肿瘤中持续激活 (3,4) ,并有致癌的潜在可能性 (5),具有抗凋亡活性 (3)。Stat3 通过 Tyr705 位点的磷酸化而激活,从而诱导二聚化、核转位、DNA 结合 (6,7)。转录激活似乎受到 MAPK 或 mTOR 通路介导的 Ser727 位点磷酸化的调控 (8,9)。Stat3 同工型的表达反应了生物学功能,Stat3α (86 kDa) 和 Stat3β (79 kDa) 相对表达水平,依赖于细胞类型、配体暴露或者细胞发育的成熟阶段 (10) 。值得注意的是,Stat3β 蛋白在羧基末端转录激活结构域缺少丝氨酸磷酸化位点 (8)。
探索与本品相关的通路。
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