Merck
CN

A5441

Sigma-Aldrich

单克隆抗 β-肌动蛋白抗体 小鼠抗

clone AC-15, ascites fluid

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别名:
单克隆抗-β-Actin
NACRES:
NA.41

生物来源

mouse

偶联物

unconjugated

抗体形式

ascites fluid

antibody product type

primary antibodies

克隆

AC-15, monoclonal

分子量

antigen 42 kDa

包含

15 mM sodium azide

species reactivity

sheep, carp, feline, chicken, rat, mouse, Hirudo medicinalis, rabbit, canine, pig, human, bovine, guinea pig

should not react with

Dictyostelium discoideum

technique(s)

immunohistochemistry (formalin-fixed, paraffin-embedded sections): suitable
indirect ELISA: suitable
indirect immunofluorescence: 1:1,000-1:2,000 using cultured human or chicken fibroblasts
western blot: 1:5,000-1:10,000 using cultured human or chicken fibroblast cell extracts

同位素/亚型

IgG1

UniProt登记号

application(s)

research pathology

运输

dry ice

储存温度

−20°C

target post-translational modification

unmodified

Gene Information

human ... ACTB(60)
mouse ... Actb(11461)
rat ... Actb(81822)

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一般描述

在鸡砂囊超薄组织冷冻切片的染色中,抗体对致密体和由结蛋白和膜相关的致密斑所占据的纵向通道连接的连续致密体进行了标记。它不会对成人的心脏和骨骼肌进行染色,除非是受到了非肌肉细胞样本的污染,或是使用了胚胎组织。
抗-β-肌动蛋白单克隆抗体(小鼠IgG1亚型)来自于免疫小鼠骨髓瘤细胞和脾细胞相互融合而产生的AC-15杂交瘤细胞。肌动蛋白是最保守的真核蛋白之一,在哺乳动物和鸟类中至少表达6种异构体。其中四种代表肌肉组织的分化标志物,而两种在几乎所有细胞中都有所发现。现有三种α-肌动蛋白(α-骨骼肌、α-心脏和 α-平滑肌),一种β-肌动蛋白(β-非肌肉)和两种γ-肌动蛋白(γ-平滑肌和γ-非肌肉)。肌动蛋白异构体显示>90%的整体序列同源性,但在其18个NH2-末端残基中只有5060%的同源性。肌动蛋白的NH2-末端区域似乎是一个主要的抗原区域,可能参与了肌动蛋白与肌球蛋白等其他蛋白的相互作用。该抗体可用于丙酮固定的冷冻切片、EM制品和显微注射实验的染色。B5、乙醇、methacam或Bouin′s溶液可用作固定剂。抗体所识别的表位对福尔马林固定和石蜡包埋具有抗性。

免疫原

略微修改的β-细胞质肌动蛋白N末端肽,Ac-Asp-Asp-Asp-Ile-Ala-Ala-Leu-Val-Ile-Asp-Asn-Gly-Ser-Gly-Lys,偶联至KLH。

应用

单克隆小鼠抗肌动蛋白抗体被用作一种上样对照,用于对来自大鼠背根神经节共培养物的免疫沉淀蛋白进行western blot分析。
单克隆小鼠抗肌动蛋白抗体被用作大鼠肝脏蛋白裂解液western blot分析的一种上样对照。
小鼠源抗-β-肌动蛋白单克隆抗体可用于:
  • 免疫荧光染色
  • 免疫印迹
  • 免疫组织化学
  • 用作蛋白芯片的对照品

生化/生理作用

肌动蛋白和肌球蛋白是许多类型细胞的组成部分,参与运动、胞质分裂、细胞质流动、分泌和吞噬等诸多细胞过程。不同物种和组织来源细胞中的肌动蛋白的免疫学和生理特性非常相似。

外形

以含有15mM叠氮化钠作为防腐剂的腹水形式提供。

其他说明

想要查看肌动蛋白抗体选择指南,请访问 www.sigmaaldrich.com/actin

免责声明

除非在我们的产品目录或产品随附的其他公司文档中另有说明,我们的产品仅供研究使用并不得用于任何其他目的,包括但不限于未经授权的商业用途、体外诊断用途、离体或体内治疗用途、任何类型的食用或应用于人或动物。

储存分类代码

10 - Combustible liquids

WGK

WGK 3

闪点(°F)

Not applicable

闪点(°C)

Not applicable

法规信息

常规特殊物品

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  1. Which document(s) contains shelf-life or expiration date information for a given product?

    If available for a given product, the recommended re-test date or the expiration date can be found on the Certificate of Analysis.

  2. What is the molecular weight of Product A5441, Monoclonal Anti-β-Actin antibody produced in mouse?

    Beta Actin has a molecular weight of 42 kDa.

  3. Can I use Product A5441, Monoclonal Anti-β-Actin antibody produced in mouse, to detect mouse beta actin?

    This antibody will detect mouse beta actin.  When used for western blotting, the secondary antibody may react with mouse IgG from the sample resulting with a bands at 50 kDa and 25 kDa (reducing conditions) or approximately 150 kDa (non-reducing conditions).  It is best to use a directly conjugated antibody for immunohistochemistry or immunofluorescent assays.  We offer Product numbers A3854, peroxidase conjugated.

  4. What IgG light chain (kappa or lambda) is associated with Clone AC-15 for Product A5441, Monoclonal Anti-β-Actin antibody produced in mouse?

    We have not determined the light chain used by monoclonal antibody AC-15.  95% of mouse antibodies have kappa light chains.

  5. Why do I have 3 bands on my western blot when using Product A5441, Monoclonal Anti-β-Actin antibody produced in mouse?

    While actin should run as a 42 kDa band when detected by Western blotting, there are recent observations that cleavage products can be seen in cells undergoing apoptosis. Initial cleavage gives a band at 41 kDa, which may appear as a doublet on the blot. In addition, further cleavage results in a major band at 30 kDa and a small band at 14 kDa which have been shown to be cleavage products due to the action of interleukin 1 -converting enzyme. This may explain the detection of smaller bands when blotting with this antibody. This banding pattern may be detected with other actin antibodies as well. Kayalar, C., et al., Proc. Natl. Acad. Sci., USA, 93:2234-2238, (1996).

  6. What is the antibody concentration of Product A5441, Monoclonal Anti-β-Actin antibody produced in mouse?

    This antibody is ascities fluid.  The amount of protein present and the amount of IgG1 present is indicated on the Lot-specific Certificate of Analysis.

  7. What peroxidase-conjugated secondary antibody should I use with Product A5441, Monoclonal Anti-β-Actin antibody produced in mouse?

    We recommend using Product No. A9044, rabbit anti-mouse IgG, peroxidase conjugate as a secondary antibody. We also offer this same antibody directly conjugated to peroxidase as Product No. A3854.

  8. How do I get lot-specific information or a Certificate of Analysis?

    The lot specific COA document can be found by entering the lot number above under the "Documents" section.

  9. How do I find price and availability?

    There are several ways to find pricing and availability for our products. Once you log onto our website, you will find the price and availability displayed on the product detail page. You can contact any of our Customer Sales and Service offices to receive a quote.  USA customers:  1-800-325-3010 or view local office numbers.

  10. What is the Department of Transportation shipping information for this product?

    Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product. 

  11. My question is not addressed here, how can I contact Technical Service for assistance?

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商品

Although actin is one of the most conserved eukaryotic proteins, it is expressed as six isoforms characterized by electrophoresis and amino acid sequence analysis. Four of the six isoforms represent differentiation markers of muscle tissues. The other two are found in practically all cells.

Loading controls in western blotting application.

Uses for immunofluorescence (IF)—where an antibody conjugated to a molecule that fluoresces when excited by lasers— include protein localization, confirmation of post-translational modification or activation, and proximity to/complexing with other proteins.

Leverage the inherent immune ability of an animal to generate antibodies that bind to specific molecules.

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实验方案

带有用于不同印迹过程工作流程步骤的Western blot实验方法,描述了蛋白从SDS聚丙烯酰胺凝胶(SDS-PAGE)到硝酸纤维素膜(NC膜)的电泳转移。也称为免疫印迹实验方法。

Western blot protocol with workflow steps for different blot procedures, describing the electrophoretic transfer of proteins from SDS polyacrylamide gels to sheets of nitrocellulose. Also known as Immunoblotting protcol.

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