Cat. # | Size | Price | Inventory |
---|---|---|---|
45332S | 100 µl (50 tests) |
REACTIVITY | H M |
SENSITIVITY | Endogenous |
MW (kDa) | |
Source/Isotype | Rabbit IgG |
Product Information
Application | Dilution |
---|---|
Immunofluorescence (Immunocytochemistry) | 1:50 - 1:200 |
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 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: 221
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
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Tyr1248 of human ErbB2 protein.
ErbB2 (HER2) 原癌基因编码了一个 185 kDa 跨膜受体样并有内在酪氨酸激酶活性的糖蛋白 (1)。虽然 ErbB2 缺乏已鉴定的配体,但在过表达时,ErbB2 激酶活性可在缺少配体的情况下被激活,也可通过杂聚肽与其他 ErbB 家族成员结合而激活 (2)。在近 40% 的人类乳腺癌中检测到 ErbB2 基因的扩增及其产物的过表达 (3)。c-Cbl 泛素连接酶在 Tyr1112 位点结合 ErbB2 会导致 ErbB2 聚泛素化,并增强这种激酶的降解 (4)。ErbB2 是治疗乳腺癌和其他癌症的一个关键治疗靶标,通过 c-Cbl 调节的蛋白水解通路靶向调节 ErbB2 降解是一种潜在治疗手段。ErbB2(与 pp60c-Src 的 Tyr416 同源)Tyr877 的激酶结构域残基的磷酸化可能参与调节 ErbB2 生物活性。ErbB2 的主要自磷酸化位点为 Tyr1248 和 Tyr1221/1222;这些位点的磷酸化使 ErbB2 偶联到 Ras-Raf-MAP 激酶信号转导通路 (1,5)。
探索与本品相关的通路。
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