Immunofluorescence analysis of beta Actin was performed using 70% confluent log phase A549 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 Actin Rabbit Polyclonal Antibody (PA121167) 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||Human, Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues A(26) P R A V F P S(33) of beta actin.|
|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 *|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/ml|
|Immunohistochemistry (Paraffin) (IHC (P))||1:100|
|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.
PA1-21167 detects beta-actin in human, mouse, rat, bovine, sheep, rabbit, feline, chicken, canine, porcine, Hirudo medicinalis (leech), Drosophila melanogaster, and guinea pig samples.
PA1-21167 has been successfully used in immunohistochemistry (FFPE) and Western blot procedures. Formalin-fixed paraffin-embedded tissue sections require boiling in 10 mM citrate buffer (pH 6.0) prior to immunostaining.
The PA1-21167 immunogen is a synthetic peptide corresponding to residues A(26) P R A V F P S(33) of beta actin.
Beta actin is one of six different actin isoforms which have been identified. Actins are highly conserved proteins that are involved in cell motility, structure and integrity.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
The Social Amoeba Dictyostelium discoideum Is Highly Resistant to Polyglutamine Aggregation.
PA1-21167 was used in western blot to identify Dictyostelium as an organism that suppress aggregation of proteins with long polyglutamine tracts
|Santarriaga S,Petersen A,Ndukwe K,Brandt A,Gerges N,Bruns Scaglione J,Scaglione KM||The Journal of biological chemistry (290:25571)||2015|
Tau deletion impairs intracellular ß-amyloid-42 clearance and leads to more extracellular plaque deposition in gene transfer models.
PA1-21167 was used in western blot to characterize Tau deletion and it leading to more extracellular plaque deposition in gene transfer models and impairment in intracellular beta-amyloid-42 clearance
|Lonskaya I,Hebron M,Chen W,Schachter J,Moussa C||Molecular neurodegeneration (9:null)||2014|
Induction of hepatic apolipoprotein A-I gene expression by the isoflavones quercetin and isoquercetrin.
PA1-21167 was used in western blot to characterize the effects of quercetin, isoquercetin, and myrescetin on apo A-I gene expression in HepG2 and Caco-2 cells
|Haas MJ,Onstead-Haas LM,Szafran-Swietlik A,Kojanian H,Davis T,Armstrong P,Wong NC,Mooradian AD||Life sciences (110:8)||2014|