Immunofluorescence analysis of alpha Tubulin was performed using 70% confluent log phase NIH/3T3 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 Rabbit Polyclonal Antibody (PA519489) at 2ug/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (A27034) at 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 (S36938). F-actin (Panel c: red) was stained with 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.
|Tested species reactivity||Bovine, Dog, Chicken, Deer, Fruit fly, Hamster, Human, Mouse, Non-human primate, Rat|
|Published species reactivity||Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide conjugated to KLH derived from within residues 400 to the C-terminus of Human alpha Tubulin.|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS, pH 7.4, with 1% BSA|
|Contains||0.02% sodium azide|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3 - 5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||1:2500|
|Western Blot (WB)||0.5 - 1 µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Immunocytochemistry (ICC)||See 1 publications below|
PA5-19489 targets alpha Tubulin in immunofluorescence, immunohistochemistry (paraffin), and Western blot applications and shows reactivity with chicken, Indian muntjac, Chinese hamster, human, fruit fly, rat, African Green Monkey, and mouse samples. Perform heat mediated antigen retrieval via the pressure cooker method before beginning immunohistochemistry staining protocol.
The PA5-19489 immunogen is synthetic peptide conjugated to KLH derived from within residues 400 to the C-terminus of Human alpha Tubulin.
PA5-19489 detects alpha Tubulin which has a predicted molecular weight of approximately 50 kDa.
Microtubules of the eukaryotic cytoskeleton perform essential and diverse functions and are composed of a heterodimer of alpha and beta tubulin. The genes encoding these microtubule constituents are part of the tubulin superfamily, which is composed of six distinct families. Genes from the alpha, beta and gamma tubulin families are found in all eukaryotes. 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 and they are highly conserved among and between species. This gene encodes an alpha tubulin that is a highly conserved homolog of a rat testis-specific alpha tubulin.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
The mesh is a network of microtubule connectors that stabilizes individual kinetochore fibers of the mitotic spindle.
PA5-19489 was used in immunocytochemistry to describe how kinetochore fibers and microtubules are associated
|Nixon FM,Gutiérrez-Caballero C,Hood FE,Booth DG,Prior IA,Royle SJ||eLife (4:null)||2015|