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抗体> 干细胞、发育与分化> Stem Cell & Lineage Markers
Vimentin Rabbit Polyclonal Antibody (KO Validated)
产品编号: AF5330
产品包装:50μl
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价格: ¥ 593.00
产品简介
使用说明
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产品编号 产品名称 产品包装 产品价格
AF5330 Vimentin Rabbit Polyclonal Antibody (KO Validated) 50μl 593.00元
来源 用途 交叉反应性 理论分子量 实际分子量
Rabbit WB, IF, IHC H, M, R 53kDa 57kDa

WB, Western blot; IP, Immunoprecipitation; IF, Immunofluorescence; IHC, Immunohistochemistry; ICC, Immunocytochemistry; FC, Flow Cytometry; ChIP, Chromatin Immunoprecipitation Assay; ChIP-seq, ChIP-sequencing.
H, Human; M, Mouse; R, Rat; C, Chicken; Cw, Cow; Dg, Dog; Gp, Guinea pig; Hm, Hamster; Hr, Horse; Mk, Monkey; Pg, Pig; Rb, Rabbit; S, Sheep; Z, Zebrafish; All, all species expected.
配套提供了Western一抗稀释液,可以用于Western检测或其它适当用途时的一抗稀释。
建议抗体使用时的稀释比例如下(实际使用时需根据抗原水平的高低作适当调整):

WB IP IF IHC ICC FC ChIP ChIP-seq
1:500-1:2000 - 1:50-1:200 1:50-1:200 1:50-1:200 - - -

抗体详细信息如下::

About this Antibody
Name Vimentin Rabbit Polyclonal Antibody (KO Validated)
Category Rabbit Polyclonal Antibody (pAb); Primary antibody
Isotype IgG
Purification method Affinity purification
Positive samples Jurkat, HeLa
Cellular location Cytoplasm
Customer validation WB (Human); IF (Human)
About the Immunogen
Immunogen Recombinant fusion protein of human Vimentin (NP_003371.2).
Sequence MSTRSVSSSSYRRMFGGPGTASRPSSSRSYVTTSTRTYSLGSALRPSTSRSLYASSPGGVYATRSSAVRLRSSVPGVRLLQDSVDFSLADAINTEFKNTRT
Gene ID 7431
Swiss Prot P08670
Synonyms VIM; CTRCT30; HEL113; vimentin
Category Stem Cell & Lineage Markers; Organelle Markers; Cytoskeletal Signaling
Background The cytoskeleton consists of three types of cytosolic fibers: microfilaments (actin filaments), intermediate filaments, and microtubules. Major types of intermediate filaments are distinguished by their cell-specific expression: cytokeratins (epithelial cells), glial fibrillary acidic protein (GFAP) (glial cells), desmin (skeletal, visceral, and certain vascular smooth muscle cells), vimentin (mesenchyme origin), and neurofilaments (neurons). GFAP and vimentin form intermediate filaments in astroglial cells and modulate their motility and shape. In particular, vimentin filaments are present at early developmental stages, while GFAP filaments are characteristic of differentiated and mature brain astrocytes. Thus, GFAP is commonly used as a marker for intracranial and intraspinal tumors arising from astrocytes. Research studies have shown that vimentin is present in sarcomas, but not carcinomas, and its expression is examined in conjunction with that of other markers to distinguish between the two. Vimentin's dynamic structural changes and spatial re-organization in response to extracellular stimuli help to coordinate various signaling pathways. Phosphorylation of vimentin at Ser56 in smooth muscle cells regulates the structural arrangement of vimentin filaments in response to serotonin. Remodeling of vimentin and other intermediate filaments is important during lymphocyte adhesion and migration through the endothelium. During mitosis, CDK1 phosphorylates vimentin at Ser56. This phosphorylation provides a PLK binding site for vimentin-PLK interaction. PLK further phosphorylates vimentin at Ser82, which might serve as memory phosphorylation site and play a regulatory role in vimentin filament disassembly. Additionally, studies using various soft-tissue sarcoma cells have shown that phosphorylation of vimentin at Ser39 by Akt1 enhances cell migration and survival, suggesting that vimentin could be a potential target for soft-tissue sarcoma targeted therapy.

包装清单:

产品编号 产品名称 包装
AF5330 Vimentin Rabbit Polyclonal Antibody (KO Validated) 50μl
AZ050 Western一抗稀释液 50ml
说明书 1份

保存条件:
Vimentin Rabbit Polyclonal Antibody (KO Validated) -20ºC保存,Western一抗稀释液-20ºC或4ºC保存,一年有效。Western一抗稀释液优先推荐4ºC保存,长期不使用可以考虑-20ºC保存,但冻融可能会导致出现轻微的浑浊和少量不溶物。
注意事项:
如果本抗体用于Western blot (WB)、免疫荧光(IF)、免疫细胞化学(ICC)等实验,请注意回收使用过的稀释抗体。回收的抗体通常至少可以重复使用5-10次。稀释后的抗体,包括已经使用过的稀释抗体,请4℃保存。
回收后重复使用的抗体,使用方法同新鲜稀释的抗体。如果在重复使用过程中发现抗体出现轻微混浊现象,可以10,000g离心1-3分钟,取上清用于后续检测。如果回收的抗体出现明显的絮状物或长霉长菌等情况,则可以考虑废弃该抗体。
提供的Western一抗稀释液也可以用于免疫荧光(IF)、免疫组化(IHC)、免疫细胞化学(ICC)等适当用途。如果希望获得最佳的检测效果,请考虑使用上述检测专用的一抗稀释液。
本产品仅限于专业人员的科学研究用,不得用于临床诊断或治疗,不得用于食品或药品,不得存放于普通住宅内。
为了您的安全和健康,请穿实验服并戴一次性手套操作。

使用说明:
请根据抗体的实际用途选择相应的使用方法。
1. Western检测:
a. 按照推荐的稀释比例用碧云天提供的Western一抗稀释液稀释抗体。
b. 把经过封闭的蛋白膜与稀释好的一抗4℃缓慢摇动过夜或室温缓慢摇动2小时,确保稀释的抗体至少能在摇动的瞬间覆盖蛋白膜。
c. 回收稀释的一抗,4℃保存以备下次继续使用。
d. 按照Western的实验步骤进行后续的洗涤、二抗孵育、洗涤和检测等操作。具体操作可以参考如下网页:
http://www.beyotime.com/western.htm
2. 免疫染色:
可以使用碧云天生产的免疫染色一抗稀释液(P0103)稀释抗体,使用后注意回收稀释好的一抗,具体操作可以参考如下网页:http://www.beyotime.com/support/immunol-staining.htm
3. 其它实验操作请自行参考适当的protocol进行。
4. 代表性图片:

Fig. 1. Western blot analysis of extracts of various cell lines, using Vimentin antibody at 1:1000 dilution. Secondary antibody: HRP-labeled Goat Anti-Rabbit IgG(H+L) (A0208) at 1:1000 dilution. Lysates/proteins: 25µg per lane. Blocking buffer: QuickBlock™ Blocking Buffer (P0231). Detection: BeyoECL Star (P0018A). Exposure time:1s. Fig. 2. Western blot analysis of extracts from normal (control) and Vimentin knockout (KO) 293T cells, using Vimentin antibody at 1:1000 dilution. Secondary antibody: HRP-labeled Goat Anti-Rabbit IgG(H+L) (A0208) at 1:1000 dilution. Lysates/proteins: 25µg per lane. Blocking buffer: QuickBlock™ Blocking Buffer (P0231). Detection: BeyoECL Star (P0018A). Exposure time:1s.
Fig. 3. Immunohistochemistry of paraffin-embedded human tonsil using Vimentin antibody at dilution of 1:150 (40x lens). Fig. 4. Immunofluorescence analysis of U2OS cells using Vimentin antibody at dilution of 1:100. Blue: DAPI for nuclear staining.

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