Cell Signaling Technology

Product Pathways - PathScan ELISA

PathScan® Total α-Tubulin Sandwich ELISA Kit #7944

cell based assay   cell-based assay   cytoskeleton   housekeeping   loading control   microtubules   normalization   sandwich ELISA   screening   total protein   tubulin  

REACTIVITY
H M Mk

No. Size Price
7944C 1 Kit ( 96 assays ) ¥6,345.00 现货查询 购买询价
Product Includes Volume Solution Color
ELISA Sample Diluent 25 ml Blue
Anti-mouse IgG, HRP-linked Antibody (ELISA Formulated) 1 ea Red (Lyophilized)
Detection Antibody Diluent 11 ml Green
HRP Diluent 11 ml Red
alpha-Tubulin Mouse Detection mAb 1 ea Green (Lyophilized)
STOP Solution #7002 11 ml Colorless
TMB Substrate #7004 11 ml Colorless
Alpha-Tubulin Rabbit Antibody Coated Microwells 96 tests
ELISA Wash Buffer (20X) 25 ml Colorless
Cell Lysis Buffer (10X) #9803 15 ml Yellowish
Sealing Tape 2 sheets

Specificity / Sensitivity

CST's PathScan® Total α-Tubulin Sandwich ELISA Kit detects endogenous levels of α-tubulin protein, as shown in Figure 1. The kit sensitivity is shown in Figure 2. Microtubule stabilizing or destabilizing agents may significantly increase or decrease the signal, respectively. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

CST's PathScan® Total β-Actin Sandwich ELISA Kit检测内源性α-tubulin蛋白,如图1所示。该试剂盒的敏感性如图2所示。微管稳定或不稳定试剂可能分别有意义的增加或减少信号。因为通过内部实验确定,该试剂盒检测已表明的物种,但是可能也会检测其它物种的同源性蛋白。

Description

The PathScan® Total α-Tubulin Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of α-tubulin. An α-tubulin rabbit antibody has been coated onto the microwells. After incubation with cell lysates, α-tubulin is captured by the coated antibody. Following extensive washing, an α-tubulin mouse detection antibody is added to detect the captured α-tubulin. An anti-mouse IgG, HRP-linked antibody is then used to recognize the bound detection antibody. HRP substrate (TMB) is added to develop color. The magnitude of the absorbance for this developed color is proportional to the quantity of α-tubulin.

PathScan® Total α-Tubulin Sandwich ELISA Kit是一个固相夹心酶联免疫吸附法,其能检测内源性α-tubulin蛋白。一个α-tubulin rabbit antibody已经被包被在微孔板上。在与细胞裂解液一起孵化后,α-tubulin蛋白被包被的抗体吸附。在大范围清洗之后,一个α-tubulin mouse detection antibody被加入去检测已被吸附的α-tubulin蛋白。然后,anti-mouse IgG, HRP-linked antibody被用于识别已结合的检测抗体。HRP底物(TMB)加入后显色。该显色的吸光度值的大小与在α-tubulin蛋白量成正比例。试剂盒里的抗体是针对该试剂盒特定制备的。

ELISA - Western correlation

ELISA - Western correlation

Figure 1. Treatment of A-431 cells with EGF stimulates phosphorylation of the EGF receptor at Tyr1086 as detected by PathScan® Phospho-EGF Receptor (Tyr1086) Sandwich ELISA Kit #7240, but does not affect the level of α-tubulin as detected by PathScan® Total α-Tubulin Sandwich ELISA Kit #7944. A-431 cells were treated with EGF (100 ng/ml, 3 min.) at 37ºC before lysis. Absorbance at 450 nm is shown in the top figure, while the corresponding western blots using Total α-Tubulin (11H10) Rabbit mAb #2125 (left) or Phospho-EGF Receptor (Tyr1086) Antibody #2220 (right) are shown in the bottom figure.

图1:使用EGF刺激EGF受体在Tyr1086位点磷酸化的A-431细胞系,随后被PathScan® Phospho-EGF Receptor (Tyr1086) Sandwich ELISA Kit #7240检测,但是通过PathScan® Total α-Tubulin Sandwich ELISA Kit #7944分析不影响α-tubulin蛋白水平。 A-431细胞在裂解之前在37ºC温度下使用EGF (100 ng/ml, 3 min.)处理。在450 nm吸光度值显示在最上图中,然而通过使用α-Tubulin (11H10) Rabbit mAb #2125 (左图)和Phospho-EGF Receptor (Tyr1086) Antibody #2220 (右图),相应的免疫印迹(western blot)显示在下图中。

Sensitivity

Sensitivity

Figure 2. The relationship between the protein concentration of the lysate from A-431 cells and the absorbance at 450 nm is shown.

图2: A-431细胞裂解的蛋白浓度与在450 nm吸光度的关系被显示。

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)。

  1. Westermann, S. and Weber, K. (2003) Nat Rev Mol Cell Biol 4, 938-47.

Application References

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Protocols

Companion Products


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

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

PathScan is a trademark of Cell Signaling Technology, Inc.

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

用户评论 --- 共 0

该产品暂无评论!

我要参与评论 :

如要参与评论请先登录网站

还没有网站账户?去注册一下吧

Products

 

Applications