Anti-alpha smooth muscle Actin 抗体 (ab5694)
製品の概要
-
製品名Anti-alpha smooth muscle Actin antibody
alpha smooth muscle Actin 一次抗体 製品一覧 -
製品の詳細Rabbit polyclonal to alpha smooth muscle Actin
-
由来種Rabbit
-
特異性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. The other actins, such as beta- and gamma-cytoplasmic, striated muscle, and myocardium are not stained by this antibody.
-
アプリケーション適用あり: IHC-FoFr, ICC, ICC/IF, WB, ELISA, IHC-P, IHC-Frmore details
-
種交差性交差種: Mouse, Rat, Chicken, Guinea pig, Cow, Dog, Human, Pig
-
免疫原
Synthetic peptide corresponding to Human alpha smooth muscle Actin. This antibody was raised against a synthetic peptide corresponding to N-terminus of actin from human smooth muscle.
-
特記事項
Actins are highly conserved proteins expressed in all eucaryotic cells. Actin filaments form part of the cytoskeleton and play essential roles in regulating cell shape and movement. Six distinct actin isotypes have been identified in mammalian cells. Each is encoded by a separated gene and is expressed in a developmentally regulated and tissue-specific manner, alpha and beta cytoplasmic actins are expressed in a wide variety of cells; whereas, expression of alpha skeletal, alpha cardiac, alpha vascular, and gamma enteric actins are more restricted to specialized muscle cell type. Smooth muscle alpha actin is of further interest because it is one of a few genes whose expression is relatively restricted to vascular smooth muscle cells. Further more, expression of smooth muscle alpha actin is regulated by hormones, cell proliferation , and altered by pathological conditions including oncogenic transformation and atherosclerosis.
Abcam recommended secondaries - Goat Anti-Rabbit HRP (ab205718) and Goat Anti-Rabbit Alexa Fluor® 488 (ab150077).
See other anti-rabbit secondary antibodies that can be used with this antibody.
製品の特性
-
製品の状態Liquid
-
保存方法Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
-
バッファーpH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS -
Concentration information loading...
-
精製度Immunogen affinity purified
-
ポリ/モノポリクローナル
-
アイソタイプIgG
-
研究分野
関連製品
-
Alternative Products
- Anti-alpha smooth muscle Actin antibody [EPR5368] (ab124964)
- Anti-alpha smooth muscle Actin antibody [E184] (Alexa Fluor® 647) (ab196919)
- Anti-alpha smooth muscle Actin antibody [E184] (Alexa Fluor® 488) (ab197240)
- Anti-alpha smooth muscle Actin antibody [EPR5368] (Alexa Fluor® 488) (ab202295)
- Anti-alpha smooth muscle Actin antibody [EPR5368] (Alexa Fluor® 555) (ab202509)
- Anti-alpha smooth muscle Actin antibody [E184] (Phycoerythrin) (ab209435)
- Anti-alpha smooth muscle Actin antibody [E184] (ab32575)
-
Compatible Secondaries
-
Isotype control
-
Positive Controls
-
Recombinant Protein
-
Related Products
- Prestained Protein Ladder - Broad molecular weight (10-245 kDa) (ab116028)
- Human alpha smooth muscle Actin peptide (ab16102)
- Phalloidin-iFluor 488 Reagent (ab176753)
- Phalloidin-iFluor 647 Reagent (ab176759)
- Anti-alpha smooth muscle Actin antibody [1A4] (Alexa Fluor® 488) (ab184675)
- Anti-alpha smooth muscle Actin antibody [1A4] (ab7817)
アプリケーション
Our Abpromise guarantee covers the use of ab5694 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
アプリケーション | Abreviews | 特記事項 |
---|---|---|
IHC-FoFr | Use at an assay dependent concentration. | |
ICC | Use at an assay dependent concentration. | |
ICC/IF | 1/100. | |
WB | Use a concentration of 0.5 - 2 µg/ml. Predicted molecular weight: 42 kDa. | |
ELISA | Use a concentration of 0.1 - 1 µg/ml. | |
IHC-P | 1/50 - 1/200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. | |
IHC-Fr | 1/200. PubMed: 18559614Fix with acetone. |
ターゲット情報
-
機能Actins are highly conserved proteins that are involved in various types of cell motility and are ubiquitously expressed in all eukaryotic cells.
-
関連疾患Defects in ACTA2 are the cause of aortic aneurysm familial thoracic type 6 (AAT6) [MIM:611788]. AATs are characterized by permanent dilation of the thoracic aorta usually due to degenerative changes in the aortic wall. They are primarily associated with a characteristic histologic appearance known as 'medial necrosis' or 'Erdheim cystic medial necrosis' in which there is degeneration and fragmentation of elastic fibers, loss of smooth muscle cells, and an accumulation of basophilic ground substance.
-
配列類似性Belongs to the actin family.
-
細胞内局在Cytoplasm > cytoskeleton.
- Information by UniProt
-
参照データベース
- Entrez Gene: 423787 Chicken
- Entrez Gene: 515610 Cow
- Entrez Gene: 477587 Dog
- Entrez Gene: 59 Human
- Entrez Gene: 11475 Mouse
- Entrez Gene: 733615 Pig
- Entrez Gene: 81633 Rat
- Omim: 102620 Human
see all -
別名
- a actin antibody
- AAT6 antibody
- ACTA_HUMAN antibody
see all
画像
-
Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin antibody (ab5694)Watcott, C.J. et al PLoS One. 2017 Aug 25;12(8):e0183871. doi: 10.1371/journal.pone.0183871. eCollection 2017
Effects of the ROCK1 inhibition on pancreatic cancer cells and cancer-associated fibroblasts
Fluorescence microscopic analysis of fasudil treated, co-cultured pancreatic cancer cells and cancer-associated fibroblasts. Cells were treated with fasudil for 48 hours and then were stained for α-SMA (red), Collagen I (green), and DNA (blue).
Alpha smooth muscle Actin (α-SMA) is detected ising ab5694 in 5% formaldehyde-fixed cells.
(From Figure 3C of Watcott et al)
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody (ab5694)Freytsis, M. et al PLoS One. 2018 Jan 5;13(1):e0190623. doi: 10.1371/journal.pone.0190623. eCollection 2018
Representative histology of aortic valve leaflets from aged mice demonstrates changes in pRb cKO AoV
A) Masson’s trichrome showing reduced collagen staining (blue) in leaflet from pRb cKO mouse with aortic regurgitation (AR). B) Movat pentachrome showing more diffuse collage staining (yellow) in fibrosa, but normal proteoglycan staining (blue) in the spongiosa layer of the leaflet from pRb cKO with AR. C) Immunohistochemistry for α-SMA, demonstrating presence of activated myofibroblasts throughout leaflets of pRb cKO mouse with and without AR. Scale bar is 50μm.
Alpha smooth muscle Actin is detected witn ab5694 at 1/1000 dilution.
(From Figure 2 of Freytsis et al)
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody (ab5694)
This picture shows formalin-fixed, paraffin embedded mouse intestine and mesentery, the optimal dilution is 1:1600 to 1:3200, incubation overnight at 4oC, counterstained with Hematoxylin.
This image was kindly supplied as part of the review by JQ Zhang.
-
Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin antibody (ab5694)This image is courtesy of an Abreview submitted by Dr Jianyuan Chai
ab5694 at 1/500 staining rat myofibroblast cells by Immunocytochemistry/ Immunofluorescence. The cells were formaldehyde fixed and blocked with 5% serum prior to incubation with the antibody for 2 hours. A FITC conjugated goat anti-rabbit IgG was used as the secondary. Nuclei were counterstained with propidium iodide.
-
Lanes 1-3 : Anti-alpha smooth muscle Actin antibody (ab5694) at 1 µg/ml
Lane 1 : HEK293 cell lysate - overexpressing alpha-Actin
Lanes 2 & 4 : 3T3 cell lysate
Lanes 3 & 5 : Mouse heart tissue homogenate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Fluor 750-conjugated goat anti-rabbit IgG (H+L) at 1/12500 dilution
Predicted band size: 42 kDa
Observed band size: 42 kDaIncubated with the primary antibody at 4°C overnight.
Incubated with the secondary antibody at room temperature for 1 hour.
-
Ab5694 positively staining smooth muscle cells in blood vessels and myoepithelial cells in the frozen tissue of cancerous human mammary gland (pink) at 1/100 dilution. Secondary: CY5 conjugated goat anti rabbit (1/100). Co immunostaining of glandular cell cytokeratin can be seen stained by FITC (green). Auto fluorescent erythrocytes that are present within blood vessels are shown (red), whilst the DAPI counter stain may clearly be seen staining nuclei (blue).
This image is courtesy of an Abreview submitted by on 22 August 2005. We do not have any further information relating to this image. -
Immunocytochemistry/ Immunofluorescence - Anti-alpha smooth muscle Actin antibody (ab5694)This image is courtesy of an anonymous Abreview
ab5694 staining Human fetal heart cells by ICC/IF. Cells were fixed with PFA and permeabilized in 0.1% saponin prior to blocking in 10% serum for 45 minutes at 37°C. The primary antibody was diluted 1/400 and incubated with the sample for 1 hour at 37°C. A Alexa Fluor® 594 conjugated goat polyclonal to rabbit IgG (H+L), diluted 1/600 was used as secondary antibody.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody (ab5694)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling alpha smooth muscle Actin with ab5694 at a dilution of 1/1000. Heat mediated antigen retrieval was performed for 35 minutes followed by cooling for 20 minutes. Sections were incubated with the primary antibody for 1 hour followed by incubation with a biotinylated secondary antibody for 30 minutes then HRP-Streptavidin for 30 minutes. Developed using DAB chromogen substrate (5-10 minutes). Counter stained with hematoxylin.
Magnification: left - 10X, right - 40X.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody (ab5694)
Immunohistochemistry (Formalin-fixed paraffin-embedded sections) analysis of skeletal muscle tissue (left) incubated with ab5694 at 1/100 at room temperature for 1 hour showing no specific staining. Right - human tonsil tissue secondary only control.
Heat mediated antigen retrieval was performed for 35 minutes followed by cooling for 20 minutes. A biotinylated secondary antibody was used for 30 minutes followed by incubation with HRP-Streptavidin for 30 minutes. Developed using DAB chromogen substrate (5-10 minutes). Counter stained with hematoxylin. Magnification 10X.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody (ab5694)Human Leiomyoma stained with ab5694.
-
All lanes : Anti-alpha smooth muscle Actin antibody (ab5694) at 1 µg/ml
Lane 1 : HeLa Nuclear
Lane 2 : HeLa whole cell
Lane 3 : A431 cell lysate
Lane 4 : Jurkat cell lysate
Lane 5 : HEK293 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Alexa Fluor anti-rabbit at 1/5000 dilution
Performed under reducing conditions.
Predicted band size: 42 kDa
Observed band size: 42 kDa
Additional bands at: 30 kDa, 35 kDa, 37 kDa, 50 kDa, 75 kDa. We are unsure as to the identity of these extra bands.
Please note that ab5694 does not appear to be specific to smooth muscle. -
All lanes : Anti-alpha smooth muscle Actin antibody (ab5694) at 1/500 dilution
Lane 1 : Rat2 myofibroblasts (untreated before treatment-0 days)
Lane 2 : Rat2 myofibroblasts (untreated for 5 days)
Lane 3 : Rat2 myofibroblasts (treated with 1ng/mL TGF beta)
Lane 4 : Rat2 myofibroblasts (treated with 10ng/mL TGF beta)
Lane 5 : Positive control (NIH3T3)
Lane 6 : Negative control (MDA-MB-469 breast carcinoma cells)
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Donkey anti rabbit (HRP) at 1/2500 dilution
Performed under reducing conditions.
Predicted band size: 42 kDaThis image is an edited version of an image submitted courtesy of an Abreview on 20 September 2005. We do not have any further information relating to this image.
-
Western blot - Anti-alpha smooth muscle Actin antibody (ab5694)This image is a courtesy of Mario TorradoAll lanes : Anti-alpha smooth muscle Actin antibody (ab5694) at 1/1000 dilution
Lanes 1-2 : Lystates prepared from pig heart tissue from normal control animals
Lanes 3-4 : Lystates prepared from pig heart tissue from experimental animals
Lysates/proteins at 4 µg per lane.
Secondary
All lanes : HRP-conjugated goat polyclonal to rabbit IgG at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 42 kDa
Observed band size: 45 kDa why is the actual band size different from the predicted?
Exposure time: 1 minute
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody (ab5694)This image is courtesy of an Anonymous Abreview
ab5694 staining alpha smooth muscle Actin in human skin tissue section by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue underwent formaldehyde fixation before heat mediated antigen retrieval in Citrate pH 6.0 and then blocked with 10% serum for 1 hour at RT. The primary antibody was diluted 1/300 and incubated with sample in 2% serum for 15 hours at 4°C. A Biotin conjugated goat polyclonal to rabbit IgG was used at dilution at 1/500 as secondary antibody.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha smooth muscle Actin antibody (ab5694)Image courtesy of an anonymous Abreview.ab5694 staining alpha smooth muscle Actin in rat lung tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).
Tissue was fixed in formaldehyde and a heat mediated antigen retrieval step was performed using citrate buffer pH 6.0. Samples were then permeabilized using 0.5 Triton X-100 for 20 minutes, blocked with 1% BSA for 30 minutes at 20°C and then incubated with ab5694 at a 1/100 dilution for 16 hours at 4°C. The secondary used was an Alexa-Fluor 568 conjugated goat anti-rabbit polyclonal used at a 1/250 dilution.
プロトコール
参考文献
This product has been referenced in:
- Gu TT et al. Pterostilbene alleviates fructose-induced renal fibrosis by suppressing TGF-ß1/TGF-ß type I receptor/Smads signaling in proximal tubular epithelial cells. Eur J Pharmacol 842:70-78 (2019). Read more (PubMed: 30336139) »
- Ambade AS et al. Triple-tyrosine kinase inhibition attenuates pulmonary arterial hypertension and neointimal formation. Transl Res 203:15-30 (2019). Read more (PubMed: 30142307) »