Immunofluorescent analysis of Actin Smooth Muscle was performed using 70% confluent log phase C2C12 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 Actin Smooth Muscle Rabbit Polyclonal Antibody (PA5-16697) 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) 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 Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoskeleton localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Bovine, Chicken, Human, Mouse, Non-human primate, Rabbit, Rat|
|Published species reactivity||Rat, Sheep, Cat, Mouse, Human, Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||A synthetic peptide from N-terminus of human alpha-smooth muscle isoform of actin|
|Storage buffer||PBS, pH 7.6, with 0.2% BSA|
|Contains||15mM sodium azide|
|Storage Conditions||4° C|
|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))||1:25|
|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 1 publications below|
|Immunohistochemistry (Frozen) (IHC (F))||See 1 publications below|
|Miscellaneous PubMed (MISC)||See 1 publications below|
|Immunohistochemistry (Paraffin) (IHC (P))||See 1 publications below|
|Immunohistochemistry (IHC)||See 11 publications below|
|Immunocytochemistry (ICC)||See 1 publications below|
PA5-16697 targets Actin Smooth Muscle in IHC (P) applications and shows reactivity with Bovine, Chicken, Human, mouse, Non-human primate, Rabbit, and Rat samples.
The PA5-16697 immunogen is a synthetic peptide from N-terminus of human alpha-smooth muscle isoform of actin.
This antibody stains smooth muscle cells in vessel walls, gut wall, and myometrium. Myoepithelial cells in breast and salivary gland are also stained. It reacts with tumors arising from smooth muscles and myoepithelial cells.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Smooth Muscle-Targeted Overexpression of Peroxisome Proliferator Activated Receptor-¿ Disrupts Vascular Wall Structure and Function.
PA5-16697 was used in western blot to learn about disruption of vascular wall structure and function by smooth muscle-targeted overexpression of peroxisome proliferator activated receptor-gamma
|Kleinhenz JM,Murphy TC,Pokutta-Paskaleva AP,Gleason RL,Lyle AN,Taylor WR,Blount MA,Cheng J,Yang Q,Sutliff RL,Hart CM||PloS one (10:null)||2015|
|Not Applicable||Not Cited||
Vascular Smooth Muscle Cells Stimulate Platelets and Facilitate Thrombus Formation through Platelet CLEC-2: Implications in Atherothrombosis.
PA5-16697 was used in immunohistochemistry - frozen section to study platelet CLEC-2 and vascular smooth muscle cells that stimulate platelets and facilitate thrombus formation and implications in atherothrombosis
|Inoue O,Hokamura K,Shirai T,Osada M,Tsukiji N,Hatakeyama K,Umemura K,Asada Y,Suzuki-Inoue K,Ozaki Y||PloS one (10:null)||2015|
PIK3CA(H1047R) induces multipotency and multi-lineage mammary tumours.
PA5-16697 was used in western blot to elucidate the potential of PIK3CA to induce multipotency during tumorigenesis in the mammary gland.
|Koren S,Reavie L,Couto JP,De Silva D,Stadler MB,Roloff T,Britschgi A,Eichlisberger T,Kohler H,Aina O,Cardiff RD,Bentires-Alj M||Nature (525:114)||2015|
|Not Applicable||Not Cited||
Neutralizing murine TGFßR2 promotes a differentiated tumor cell phenotype and inhibits pancreatic cancer metastasis.
PA5-16697 was used in immunohistochemistry - paraffin section to analyze promotion of a differentiated tumor cell phenotype and inhibition of pancreatic cancer metastasis by neutralizing murine TGF-betaR2
|Ostapoff KT,Cenik BK,Wang M,Ye R,Xu X,Nugent D,Hagopian MM,Topalovski M,Rivera LB,Carroll KD,Brekken RA||Cancer research (74:4996)||2014|
Solitary tumours associated with Jaagsiekte retrovirus in sheep are heterogeneous and contain cells expressing markers identifying progenitor cells in lung repair.
PA5-16697 was used in immunohistochemistry to study the heterogeneity and the presence of lung repair progenitor cells in Jaagsiekte retrovirus-associated solitary tumors in sheep
|De las Heras M,de Martino A,Borobia M,Ortín A,Álvarez R,Borderías L,Giménez-Más JA||Journal of comparative pathology (150:138)||2014|
Compartmental and temporal dynamics of chronic inflammation and airway remodelling in a chronic asthma mouse model.
PA5-16697 was used in immunohistochemistry to study airway remodeling dynamics and the effects of inhaled corticosteroid therapy in a murine model of asthma
|Alrifai M,Marsh LM,Dicke T,K¿l¿ç A,Conrad ML,Renz H,Garn H||PloS one (9:null)||2014|
The cAMP effector EPAC activates Elk1 transcription factor in prostate smooth muscle, and is a minor regulator of ¿1-adrenergic contraction.
PA5-16697 was used in immunohistochemistry to study the modulation of the Elk1 transcription factor by EPAC in prostate smooth muscle
|Hennenberg M,Strittmatter F,Schmetkamp H,Rutz B,Walther S,Stief CG,Gratzke C||Journal of biomedical science (20:null)||2013|
The impact of KRAS mutations on VEGF-A production and tumour vascular network.
PA5-16697 was used in immunohistochemistry to study the differential effects on the production of VEGF-A and tumor vasculature development of codon 12 and codon 13 KRAS mutations
|Figueras A,Arbos MA,Quiles MT,Viñals F,Germà JR,Capellà G||BMC cancer (13:null)||2013|
Pericyte coverage of differentiated vessels inside tumor vasculature is an independent unfavorable prognostic factor for patients with clear cell renal cell carcinoma.
PA5-16697 was used in immunohistochemistry to study the prognostic significance of pericyte coverage of differentiated tumor microvessels in clear cell renal cell carcinoma
|Cao Y,Zhang ZL,Zhou M,Elson P,Rini B,Aydin H,Feenstra K,Tan MH,Berghuis B,Tabbey R,Resau JH,Zhou FJ,Teh BT,Qian CN||Cancer (119:313)||2013|
Recruitment of podoplanin positive cancer-associated fibroblasts in metastatic lymph nodes predicts poor prognosis in pathological N2 stage III lung adenocarcinoma.
PA5-16697 was used in immunohistochemistry to study the prognostic significance podoplanin-expressing positive cancer-associated fibroblasts in patients with pathological N2 stage III lung adenocarcinoma
|Neri S,Ishii G,Taira T,Hishida T,Yoshida J,Nishimura M,Nagai K,Ochiai A||Annals of surgical oncology (19:3953)||2012|
Adenosine A(2a) receptor stimulation prevents hepatocyte lipotoxicity and non-alcoholic steatohepatitis (NASH) in rats.
PA5-16697 was used in immunohistochemistry to study the molecular mechanisms underlying the protection against lipotoxicity and non-hepatic steatosis of adenosine A2a receptor activation
|Imarisio C,Alchera E,Sutti S,Valente G,Boccafoschi F,Albano E,Carini R||Clinical science (London, England : 1979) (123:323)||2012|
Expression of NOS and VEGF in feline mammary tumours and their correlation with angiogenesis.
PA5-16697 was used in immunohistochemistry to study the expression of eNOS, iNOS and VEGF and their correlation with angiogenesis in feline mammary tumors
|Islam MS,Matsumoto M,Hidaka R,Miyoshi N,Yasuda N||Veterinary journal (London, England : 1997) (192:338)||2012|
Deleted in colorectal carcinoma suppresses metastasis in p53-deficient mammary tumours.
PA5-16697 was used in immunohistochemistry to study the suppression of p53-deficient mammary tumor metastasis by DCC
|Krimpenfort P,Song JY,Proost N,Zevenhoven J,Jonkers J,Berns A||Nature (482:538)||2012|
Generation and hepatic differentiation of human iPS cells.
PA5-16697 was used in immunohistochemistry to develop a method for the generation and hepatic differentiation of human induced pluripotent stem cells
|Ishikawa T,Hagiwara K,Ochiya T||Methods in molecular biology (Clifton, N.J.) (826:103)||2011|
Luminal expression of PIK3CA mutant H1047R in the mammary gland induces heterogeneous tumors.
PA5-16697 was used in immunohistochemistry to elucidate the contribution of PIK3CA H1047R to breast cancer pathogenesis.
|Meyer DS,Brinkhaus H,Müller U,Müller M,Cardiff RD,Bentires-Alj M||Cancer research (71:4344)||2011|
Giant cell tumor of bonelike lesion in a Trp53 mutant mouse.
PA5-16697 was used in immunocytochemistry and immunohistochemistry to report on a case of giant cell tumor of bonelike lesion in a Trp53 mutant mouse
|Radaelli E,Rustighi A,Scanziani E||Toxicologic pathology (40:675)||2012|