alpha Tubulin Monoclonal Antibody (DM1A)/α-Tubulin内参抗体

货号：62204

规格：100 µL

价格：2753

产品类型：内参抗体

品牌：Thermo Fisher

物种：人/小鼠/大鼠

宿主：小鼠

抗体亚型：其它

荧光染料：其它

点击Invitrogen内参抗体集，查看更多内参抗体

抗体类型:	单抗	同型对照：	IgG1, kappa
免疫原性：	Native chick brain microtubules	用法：	2 µg/mL（IP）；2 µg/mL（IF）；1 µg/mL(WB);1:400(IHC(P))

Product Specific Information62204 detects alpha tubulin in human, mouse, rat, bovine, porcine, chicken, amphibian, gerbil and guinea pig samples.62204 has been successfully used in Western blot, immunohistochemistry, immunoprecipitation, electron microscopy, immunocytochemistry and immunofluorescent applications.The 62204 immunogen is native chick brain microtubules. This antibody recognizes an epitope within amino acids 426-450 of alpha tubulin.Background/Target InformationAlpha Tubulin is a part of a heterodimer consisting of alpha and beta tubulin subunits. The alpha and beta tubulins, which are each about 55 kDa MW, are homologous but not identical. Alpha-beta tubulin heterodimer is the basic building block of microtubules, and this intracellular cylindrical filamentous structure is present in almost eukaryotic cells. Microtubules serve as structural supports and lines of transport within the cell, as well as serving a key role in mitosis. Microtubules of the eukaryotic cytoskeleton perform essential and diverse functions and are composed of a heterodimer of alpha and beta tubulins. The alpha and beta tubulins represent the major components of microtubules, while gamma tubulin plays a critical role in the nucleation of microtubule assembly. There are multiple alpha and beta tubulin genes, which are highly conserved among species. The gene that encodes alpha tubulin is highly similar to the mouse and rat Tuba1 gene. Northern blotting studies have shown that the gene expression is predominantly found in morphologically differentiated neurologic cells.For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

数据

alpha Tubulin Antibody (62204) in IFImmunofluorescent analysis of Alpha Tubulin was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Alpha Tubulin (DM1A) Mouse Monoclonal Antibody (Product # 62204) at 2 µg/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification. alpha Tubulin Antibody (62204) in WBWestern blot analysis was performed on membrane enriched extract (30 µg) of SH-SY5Y (Lane 1), U-87 MG (Lane 2) COLO 205 (Lane 3), HT-29 (Lane 4), PC-3 (Lane 5), MDA-MB-231 (Lane 6), MCF7 (Lane 7), NIH/3T3 (Lane 8), A-431 (Lane9) and Jurkat (Lane10). The blots were probed with Anti-Alpha Tubulin Mouse Monoclonal Antibody (Product # 62204, 2 µg/mL) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.4 µg/mL, 1:2500 dilution). A ~ 55 kDa band corresponding to Alpha Tubulin was observed across cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE®12 % Bis-Tris gel (Product # NP0342BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody using iBind™ Flex Western Starter Kit (Product # SLF2000S). Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106). alpha Tubulin Antibody (62204) in WBWestern blot analysis was performed on whole cell extracts (30µg lysate) of A-431 (Lane 1), COS-7 (Lane 2), MDCK (Lane 3), NIH/3T3 (Lane 4), HT-29 (Lane 5), PC-12 (Lane 6), Neuro-2a (Lane 7), tissue extracts of Mouse Pancreas (Lane 8) and Rat Brain (Lane 9). The blot was probed with Anti- alpha Tubulin Mouse Monoclonal Antibody (Product # 62204, 1µg/mL) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25µg/mL, 1:4000 dilution). A 55 kDa band corresponding to alpha Tubulin was observed across the cell lines and tissues tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 4-12 % Bis-Tris gel (Product # NP0322BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).

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参考文献：

1.Molecular neurobiologyThe Motor KIF5C Links the Requirements of Stable Microtubules and IGF-1 Receptor Membrane Insertion for Neuronal Polarization."62204 was used in western blot to provide a mechanistic link between three early polarity events needed for neuronal polarity"AuthorsOksdath M,Guil AFN,Grassi D,Sosa LJ,Quiroga S2.Scientific reportsThe PTEN phosphatase functions cooperatively with the Fanconi anemia proteins in DNA crosslink repair."62204 was used in western blot to investigate the role of the PTEN phosphatase in DNA interstrand crosslink repair"AuthorsVuono EA,Mukherjee A,Vierra DA,Adroved MM,Hodson C,Deans AJ,Howlett NG3.PloS oneMPT0B098, a Microtubule Inhibitor, Suppresses JAK2/STAT3 Signaling Pathway through Modulation of SOCS3 Stability in Oral Squamous Cell Carcinoma."62204 was used in immunocytochemistry and western blot to investigate the effect of a microtubule inhibitor, MPT0B098, on STAT3 signaling in oral squamous cell carcinoma"AuthorsPeng HY,Cheng YC,Hsu YM,Wu GH,Kuo CC,Liou JP,Chang JY,Jin SL,Shiah SG4.Scientific reportsPurifying Cytokinetic Cells from an Asynchronous Population."62204 was used in immunocytochemistry and western blot to assess and asynchronous population to purify cytokinetic cells"AuthorsPanet E,Ozer E,Mashriki T,Lazar I,Itzkovich D,Tzur A5.PloS oneA second tubulin binding site on the kinesin-13 motor head domain is important during mitosis."62204 was used in immunocytochemistry to study the mitotic importance of the second tubulin binding site in the kinesin13 motor head domain"AuthorsZhang D,Asenjo AB,Greenbaum M,Xie L,Sharp DJ,Sosa H

技术参数

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