用户验证的应用
经验证应用
| 项目 | 详情 |
|---|---|
| WB | Western blot analysis of extracts from RAW264.7 (lane 2(20μg), SH-SY5Y (lane 3(20μg), Neuro-2a (lane 4(20μg), Jurkat (lane 5(20μg) using beta III Tubulin Antibody. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in TBST for 2 hour at room temperature. The primary antibody (1/1000) and Loading control antibody (Beta Actin, HY-P80438, 1/3000) was used in 5% BSA in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (HY-P8004/HY-P8001, 1/10,000) was used for 1 hour at room temperature. |
| ICC | Immunocytochemistry analysis of HEK293 cells labeling beta III Tubulin Antibody ( HY-P84546 ) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with BSA for Immunol Staining for 10 min at room temperature. Cells were then incubated with beta III Tubulin Antibody ( HY-P84546 ) at 1/50 dilution in BSA for Immunol Staining at 4 ℃ overnight. AF488-conjugated AffiniPure Goat Anti-Mouse IgG H&L（HY-P8005, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue). Immunocytochemistry analysis of Neuro-2a cells labeling beta III Tubulin Antibody ( HY-P84546 ) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with BSA for Immunol Staining for 10 min at room temperature. Cells were then incubated with beta III Tubulin Antibody ( HY-P84546 ) at 1/50 dilution in BSA for Immunol Staining at 4 ℃ overnight. AF488-conjugated AffiniPure Goat Anti-Mouse IgG H&L（HY-P8005, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue). |
| FC | Flow Cytometry analysis of MCF-7 cells labelling beta III Tubulin (red) with beta III Tubulin Antibody (anti-beta III Tubulin) ( HY-P84546 ). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. Then cells were stained with the primary antibody at 1/200 dilution for an hour at 4℃. Goat Anti-Mouse IgG H&L (AF488) ( HY-P8005 ) was used as the secondary antibody at 1/1,000 dilution for 30 minutes at 4℃. Mouse IgG Isotype Control (HY-P807578, blue) was used as the isotype control, cells without incubation with primary antibody were used as the unlabeled control (black). |
推荐稀释比例
- WB1:500-1:2000
- ICC/IF1:200-1:1000
- FC1:200-1:400
- ELISA1:10000
详细信息
背景信息
功能： βIII 微管蛋白 (TUBB3) 属于微管蛋白家族，微管蛋白是微管的主要组成成分，微管是由 α-和 β-微管蛋白异二聚体组成的蛋白丝。微管通过在其末端添加 GTP 结合型微管蛋白二聚体实现生长，末端会形成一个稳定帽。稳定帽下方的 α-β 微管蛋白异二聚体处于 GDP 结合状态，这是由 α-微管蛋白的 GTP 酶活性导致的。TUBB3 在轴突的正确导向和维持中发挥关键作用。NTN1/Netrin-1 与其受体 UNC5C 结合后，可能会使 UNC5C 与微管中聚合的 TUBB3 解离，从而提高微管动态性，导致轴突排斥。在背根神经节轴突向脊髓的投射过程中发挥作用 [1] [2] [3] [4] [5] 。 亚细胞定位： 细胞质、细胞骨架；细胞突起、生长锥；细胞突起、片状伪足；细胞突起、丝状伪足 表达水平： 组织特异性：该基因主要表达于中枢和周围神经系统，在大多数癌变组织中表达显著增强。 异构体 & 翻译后修饰： Q13509 有两种异构体：Q13509-1：50433 Da (预测值)；Q13509-2：42433 Da (预测值)。 C 端的一些谷氨酸残基发生多聚谷氨酰化，导致 γ-羧基上形成多聚谷氨酸链 (PubMed:26875866)。多聚谷氨酰化在微管蛋白 (SPAST) 切割微管的过程中起着关键作用。SPAST 优先识别并作用于带有短多聚谷氨酸尾的微管：SPAST 的切割活性随着每个微管蛋白上的谷氨酸数量从一个增加到八个而增强，但超过此谷氨酰化阈值后则下降 (PubMed:26875866)。谷氨酰化也参与纤毛运动 (基于相似性)；由于人类缺乏功能性 TTLL10，C 端的一些谷氨酸残基发生单甘氨酰化而非多甘氨酰化。单甘氨酰化主要限于整合到纤毛和鞭毛轴丝中的微管蛋白，这对于它们的稳定性和维持至关重要。鞭毛的甘氨酰化控制精子运动。多谷氨酰化和单甘氨酰化可以共存于同一蛋白质的相邻残基上，降低甘氨酰化水平会增加多谷氨酰化水平，反之亦然；在细胞周期中，从中期到末期，CDK1 会在 Ser-172 位点对其进行磷酸化，但在间期则不会。这种磷酸化会抑制微管蛋白整合到微管中。 亚基： α-和β-微管蛋白异二聚体 (PubMed:34996871, PubMed:35482892, PubMed:38305685, PubMed:38609661)。