Cell Signaling Technology

Product Pathways - Cytoskeletal Signaling

Acetyl-α-Tubulin (Lys40) (D20G3) XP® Rabbit mAb #5335

No. Size Price
5335S 100 µl ( 10 western blots ) ¥4,050.00 现货查询 购买询价
5335T 20 µl ( 2 western blots ) ¥1,600.00 现货查询 购买询价
5335 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat,Monkey,Zebrafish, Endogenous 52 Rabbit IgG
IP 1:50
F 1:200
IF-IC 1:800

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, F=Flow Cytometry, IF-IC=Immunofluorescence (Immunocytochemistry),

Homology

Species predicted to react based on 100% sequence homology: Xenopus,

Specificity / Sensitivity

Acetyl-α-Tubulin (Lys40) (D20G3) XP® Rabbit mAb detects endogenous levels of α-tubulin only when acetylated at Lys40. This amino acid is not conserved in β-tubulin.

Acetyl-α-Tubulin (Lys40) (D20G3) XP® Rabbit mAb兔单抗检测仅在Lys40位点乙酰化的内源性tubulin蛋白。该氨基酸不在β-tubulin蛋白中保守。

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic acetylpeptide corresponding to residues surrounding Lys40 of human α-tubulin.

通过人工合成人源α-tubulin蛋白Lys40位点周围相应的乙酰化片段去免疫动物从而制备出单克隆抗体。

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, untreated or TSA-treated (400 nM for 16 hours), using Acetyl-α-Tubulin (Lys40) (D20G3) XP® Rabbit mAb (upper) and α-Tubulin (11H10) Rabbit mAb #2125 (lower). The acetyl-specificity of the antibody was verified by blocking with an acetyl- or non-acetylpeptide.

使用Acetyl-α-Tubulin (Lys40) (D20G3) XP® Rabbit mAb (上图)和α-Tubulin (11H10) Rabbit mAb #2125 (下图),免疫印迹(Western Blot)分析HeLa细胞中Acetyl-α-Tubulin (Lys40)和α-Tubulin (11H10)蛋白水平。细胞分为untreated或TSA-treated (400 nM for 16 hours)。

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells, untreated (left) or treated with Trichostatin A #9950 (right), using Acetyl-α-Tubulin (Lys40) (D20G3) XP® Rabbit mAb (green). Actin filaments have been labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

使用Acetyl-α-Tubulin (Lys40) (D20G3) XP® Rabbit mAb (绿色)标记,共聚焦免疫荧光分析HeLa细胞,细胞分为untreated (左图)或treated with Trichostatin A #9950 (右图)。DY-554 phalloidin标记微丝蛋白(红色)。蓝色伪彩= DRAQ5® #4084 (DNA荧光染料)。

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of HeLa cells, untreated (blue) or treated with Trichostatin A (TSA) #9950 (1uM, 16 hours) using Acetyl-alpha-Tubulin (Lys40) (D20G3) XP® Rabbit mAb. Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor 488 Conjugate) #4412 was used as a secondary antibody.

Background

The cytoskeleton consists of three types of cytosolic fibers: microtubules, microfilaments (actin filaments), and intermediate filaments. Globular tubulin subunits comprise the microtubule building block, with α/β-tubulin heterodimers forming the tubulin subunit common to all eukaryotic cells. γ-tubulin is required to nucleate polymerization of tubulin subunits to form microtubule polymers. Many cell movements are mediated by microtubule action, including the beating of cilia and flagella, cytoplasmic transport of membrane vesicles, chromosome alignment during meiosis/mitosis, and nerve-cell axon migration. These movements result from competitive microtubule polymerization and depolymerization or through the actions of microtubule motor proteins (1).

细胞骨架由三种类型的细胞质基质纤维组成:微管、微丝(肌动蛋白纤维)和中间纤维。球形微管蛋白亚基包含微管建筑块,由α/β-tubulin异源二聚体形成所有真核细胞的tubulin亚单位。γ-tubulin被需要在tubulin亚单位的成核聚合作用中去形成微管聚合物。许多细胞运动是通过微管动力介导的,包括纤毛和鞭毛的跳动、膜泡的核质转运、在减数分裂/有丝分裂期间染色体排布和神经轴突的迁移。这些运动由于竞争性微管聚合作用和解聚作用或通过微管动力蛋白的运动(1)。

The Elongator complex catalytic subunit (Elp3) acetylates α-tubulin at Lys40 while the histone deacetylase HDAC6 functions as a tubulin deacetylase. This post-transcriptional modification may be required for dynamic cell shape remodeling, cell motility, tubulin stability and terminal branching of cortical neurons (2,3).

elongator 复合物的催化亚基(Elp3)使α-tubulin蛋白在Lys40位点乙酰化,然而组蛋白脱乙酰酶HDAC6的功能是tubulin蛋白的脱乙酰酶。这个转录后修饰可能对动态的细胞形态的重塑、细胞能动性、tubulin稳定性和皮层神经元的终端分支是需要的(2-3)。

  1. Westermann, S. and Weber, K. (2003) Nat Rev Mol Cell Biol 4, 938-47.
  2. Creppe, C. et al. (2009) Cell 136, 551-564.
  3. Hubbert, C. et al. (2002) Nature 417, 455-458.

Application References

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For Research Use Only. Not For Use In Diagnostic Procedures.

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Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

XP is a registered trademark of Cell Signaling Technology, Inc.

Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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