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 (PA116947) 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, Hamster, Human, Mouse, Non-human primate, Pig, Rat, Xenopus, Zebrafish|
|Published species reactivity||Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide between amino acids 1-100 of the human Beta-Tubulin protein.|
|Purification||Ammonium sulfate precipitation|
|Contains||0.02% sodium azide|
|Storage Conditions||Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.|
|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))||1:1000-1:2000|
|Western Blot (WB)||1:1,000 - 1:5,000|
* 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.
Suggested positive control: antigen standard for TUBB (transient overexpression lysate), Hela whole cell extract.
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. Antibodies against beta Tubulin are useful as loading controls for Western Blotting.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
The Role of Sonic Hedgehog in the Specification of Human Cortical Progenitors In Vitro.
PA1-16947 was used in western blot to study the relation between human cortical formation and impaired sonic hedgehog signaling
|Radonji¿ NV,Memi F,Ortega JA,Glidden N,Zhan H,Zecevic N||Cerebral cortex (New York, N.Y. : 1991) (26:131)||2016|
The new 4-O-methylhonokiol analog GS12021 inhibits inflammation and macrophage chemotaxis: role of AMP-activated protein kinase ¿ activation.
PA1-16947 was used in western blot to determine if methylhonokiol analogs inhibit the expression of inflammatory genes in macrophages and adipocytes
|Kim S,Ka SO,Lee Y,Park BH,Fei X,Jung JK,Seo SY,Bae EJ||PloS one (10:null)||2015|
Inhibition of adipocyte inflammation and macrophage chemotaxis by butein.
PA1-16947 was used in western blot to study the mechanisms underlying the inhibitory effects of butein on adipocyte inflammation and macrophage chemotaxis
|Wang Z,Lee Y,Eun JS,Bae EJ||European journal of pharmacology (738:40)||2014|
Synthesis of the Proposed Structure of Damaurone D and Evaluation of Its Anti-inflammatory Activity.
PA1-16947 was used in western blot to describe a method to synthesize damaurone D and test its immunomodulatory activity
|Han YT,Wang Z,Bae EJ||Chemical and pharmaceutical bulletin (63:907)||2015|