Immunofluorescence analysis of beta Tubulin was performed using 70% confluent log phase SH-SY5Y 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 Beta Tubulin Rabbit Polyclonal Antibody (PA121153) 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||Many|
|Published species reactivity||Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues N(416) D L V S E Y Q Q Y Q D A T A(430) of human beta Tubulin.|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS, pH 7.4, with 1% BSA|
|Contains||0.09% sodium azide|
|Storage Conditions||4° C, do not freeze|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Functional Assay (FN)||Assay-dependent|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||1:200|
|Immunoprecipitation (IP)||Assay dependent|
|Western Blot (WB)||1 - 3 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.
|Western Blot (WB)||See 2 publications below|
PA1-21153 detects beta-Tubulin in guinea pig, human, mouse, rat, fungus, S. cerevisiae, bovine, porcine, sea urchin, gerbil, and chicken samples.
PA1-21153 has been successfully used in immunohistochemistry (paraffin), immunoprecipitation and Western blot procedures. Suggested positive controls for this product are MAD109 cells, skin and lung.
The PA1-21153 immunogen is a synthetic peptide corresponding to residues N(416) D L V S E Y Q Q Y Q D A T A(430) of human beta Tubulin.
Tubulin is the major building block of microtubules. This intracellular cylindrical filamentous structure is present in almost all eukaryotic cells. Microtubules function as structural and mobile elements in mitosis, intracellular transport, flagellar movement, and in the cytoskeleton.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Hypoxia Signaling Cascade for Erythropoietin Production in Hepatocytes.
PA1-21153 was used in western blot to determine the regulatory mechanisms of hepatic Epo production
|Tojo Y,Sekine H,Hirano I,Pan X,Souma T,Tsujita T,Kawaguchi S,Takeda N,Takeda K,Fong GH,Dan T,Ichinose M,Miyata T,Yamamoto M,Suzuki N||Molecular and cellular biology (35:2658)||2015|
mTORC1 Down-Regulates Cyclin-Dependent Kinase 8 (CDK8) and Cyclin C (CycC).
PA1-21153 was used in western blot to demonstrate that mTORC1 activation downregulates the CDK8-CycC complex and elevates lipogenic protein expression.
|Feng D,Youn DY,Zhao X,Gao Y,Quinn WJ,Xiaoli AM,Sun Y,Birnbaum MJ,Pessin JE,Yang F||PloS one (10:null)||2015|