Cat. # | Size | Price | Inventory |
---|---|---|---|
52953S | 100 µl |
REACTIVITY | H |
SENSITIVITY | Endogenous |
MW (kDa) | 180-250, 70 |
Source/Isotype | Rabbit IgG |
Product Information
Application | Dilution |
---|---|
Western Blotting | 1:1000 |
Immunofluorescence (Immunocytochemistry) | 1:1600 - 1:3200 |
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
Achieve higher quality immunofluorescent images using the efficient and cost-effective, pre-made reagents in our #12727 Immunofluorescence Application Solutions Kit
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade 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.
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: 24
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
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro178 of human syndecan 1 protein.
Syndecans 是一个 1 型跨膜硫酸乙酰肝素蛋白多糖家族,在哺乳动物中由四个成员组成 (SDC1-4) (1),由 4 个 Syndecan 基因编码。Syndecans 参与胚胎发育、肿瘤发生和血管生成 (2)。细胞外结构域含有硫酸乙酰肝素和硫酸软骨素链的结合位点,有助于与一系列蛋白相互作用,包括大量生长因子。此外,疏水性的 C 末端胞内结构域可以和含有 PDZ 结构域的蛋白结合 (2)。这些相互作用使 Syndecans 可以作为细胞膜信号的重要整合因子 (3)。Syndecans 经过蛋白裂解,可使其胞外结构域释放(脱落),将膜结合蛋白转化为可溶性分子效应器 (4)。
在血液系统疾病中,Syndecan 1 (SDC1) 是浆细胞分化的一个特异性标记物 (5-7)。这种细胞表面蛋白多糖在正常上皮细胞和组织以及不同类型的癌组织中也有表达 (8-11)。The extracellular shed form of syndecan 1 remains soluble or accumulates in the extracellular matrix, where it binds growth factors, cytokines, and other extracellular matrix proteins (12,13). 该结合可激活结合的生长因子或细胞因子的信号转导,促进肿瘤生长、播散、血管生成和骨质溶解 (14-17)。因此,多聚糖 1 蛋白及其脱落形式的水平可以充当一系列恶性肿瘤的预后因素 (6,18,19)。最近发现 Syndecan 1 是胰腺癌中大蛋白细胞增多症的关键介质 (20)。
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