Anti-alpha smooth muscle Actin 抗体 [EPR5368] (ab124964)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR5368] to alpha smooth muscle Actin
- Suitable for: WB, IHC-P, ICC/IF, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
製品の概要
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製品名
Anti-alpha smooth muscle Actin antibody [EPR5368]
alpha smooth muscle Actin 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPR5368] to alpha smooth muscle Actin -
由来種
Rabbit -
アプリケーション
適用あり: WB, IHC-P, ICC/IF, Flow Cyt (Intra)more details -
種交差性
交差種: Mouse, Rat, Human -
免疫原
Synthetic peptide within Human alpha smooth muscle Actin aa 1-100 (N terminal). The exact sequence is proprietary.
Database link: P62736 -
ポジティブ・コントロール
- WB: HeLa, HEK-293, U937, SV40LT-SMC, A549, C2C12, A431 and NIH/3T3 cell lysates. Mouse and rat brain and heart tissue lysates. Human heart, skeletal muscle and lung tissue lysates. IHC-P: Human prostatic carcinoma, stomach carcinoma, tonsil, heart, skeletal muscle (exhibits vascular smooth muscle staining), normal stomach, liver, colon, tonsil and ovary tissues; Mouse Uterus and stomach tissue. ICC/IF: A-673 and HeLa cells. Flow Cyt (intra): Jurkat cells.
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特記事項
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
製品の特性
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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. Avoid freeze / thaw cycle. -
解離定数(KD 値)
KD = 2.20 x 10 -11 M Learn more about KD -
バッファー
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, PBS -
Concentration information loading...
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精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
EPR5368 -
アイソタイプ
IgG -
研究分野
関連製品
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Alternative Versions
- Alexa Fluor® 488 Anti-alpha smooth muscle Actin antibody [EPR5368] (ab202295)
- Alexa Fluor® 647 Anti-alpha smooth muscle Actin antibody [EPR5368] (ab202296)
- Alexa Fluor® 555 Anti-alpha smooth muscle Actin antibody [EPR5368] (ab202509)
- Alexa Fluor® 594 Anti-alpha smooth muscle Actin antibody [EPR5368] (ab202510)
- PE Anti-alpha smooth muscle Actin antibody [EPR5368] (ab208844)
- Anti-alpha smooth muscle Actin antibody [EPR5368] - BSA and Azide free (ab220795)
- Anti-Vimentin antibody [LN-6] (ab230171)
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
- U937 cytoplasmic extract lysate (ab14904)
- A-431 whole cell lysate (ab30132)
- Mouse brain tissue lysate - total protein (ab30151)
- Mouse heart normal tissue lysate - total protein (ab30291)
- Mouse brain tissue lysate - total protein (0 days) (ab7188)
- Mouse brain tissue lysate - total protein (14 days) (ab7189)
- Mouse brain tissue lysate - total protein (7 days) (ab7190)
- Mouse heart tissue lysate - total protein (0 days) (ab7193)
- Mouse heart tissue lysate - total protein (14 days) (ab7194)
- HEK-293 whole cell lysate (ab7902)
- A-431 whole cell lysate (ab7909)
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Recombinant Protein
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Related Products
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab124964の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
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WB | (1) |
1/10000 - 1/50000. Detects a band of approximately 42 kDa (predicted molecular weight: 42 kDa).
For unpurified use at 1/1000. |
IHC-P | (10) |
1/1000 - 1/2500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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ICC/IF | (3) |
1/250 - 1/500.
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Flow Cyt (Intra) |
1/10 - 1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
特記事項 |
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WB
1/10000 - 1/50000. Detects a band of approximately 42 kDa (predicted molecular weight: 42 kDa). For unpurified use at 1/1000. |
IHC-P
1/1000 - 1/2500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
ICC/IF
1/250 - 1/500. |
Flow Cyt (Intra)
1/10 - 1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ターゲット情報
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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
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参照データベース
- Entrez Gene: 59 Human
- Entrez Gene: 11475 Mouse
- Entrez Gene: 81633 Rat
- Omim: 102620 Human
- SwissProt: P62736 Human
- SwissProt: P62737 Mouse
- SwissProt: P62738 Rat
- Unigene: 500483 Human
see all -
別名
- a actin antibody
- AAT6 antibody
- ACTA_HUMAN antibody
see all
画像
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Immunohistochemical analysis of formalin-fixed paraffin-embedded human endometrium labelling alpha smooth muscle actin with ab124964 at a concentration of 0.07µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was conducted for 32min with ULTRA cell conditioning solution (CC1 pH8.5). ab124964 anti alpha smooth muscle actin antibody was incubated at 37°C for 16min. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.
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Tissue Microarrays stained for Anti-alpha smooth muscle Actin antibody [EPR5368] using ab124964 in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negaive (cross mark) staining per sample type tested. The section was incubated with ab124964 for 30 mins at room temperature followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.
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ab124964 staining alpha smooth muscle Actin in SV40LT-SMC cells (positive control, top panel) and A431 cells (negative control, bottom panel). The cells were fixed with 100% methanol (5 min) then permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab124964 at 0.5μg/ml concentration and ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8). -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human stomach tissue labelling alpha smooth muscl Actin with purified ab124964 at 1/1000. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with Hematoxylin.
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ab124964 staining alpha smooth muscle Actin in wild-type HeLa cells (top panel) and ACTA2 knockout HeLa cells (bottom panel). The cells were fixed with 100% methanol (5 min) then permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab124964 at 1/500 dilution and ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8). -
All lanes : Anti-alpha smooth muscle Actin antibody [EPR5368] (ab124964) at 1/10000 dilution
Lane 1 : Wild-type HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : ACTA2 knockout HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 42 kDa
Observed band size: 42 kDaLanes 1 - 2: Merged signal (red and green). Green - ab124964 observed at 42 kDa. Red - loading control, ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.
ab124964 was shown to react with alpha smooth muscle Actin in wild-type HeLa cells in western blot. Loss of signal was observed when ACTA2 knockout sample was used. Wild-type HeLa and ACTA2 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab124964 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4°C at a 1 in 10000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Intracellular Flow Cytometry analysis of Jurkat (human T cell leukemia cell line from peripheral blood) cells labelling alpha smooth muscle Actin with purified ab124964 at 1/30 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Green - Isotype control, rabbit monoclonal IgG.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse stomach tissue labelling alpha smooth muscl Actin with ab124964 at 1/1000. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Counterstained with Hematoxylin.
Positive staining on smooth muscle in mouse stomach.
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Immunocytochemistry/Immunofluorescence analysis of HeLa cells labeling alpha smooth muscle Actin with ab124964 at 1/500. Cells were fixed with 4% paraformaldehyde and permeabilised with 0.1% tritonX-100. An Goat anti rabbit IgG(Alexa Fluor® 488)
ab150077 (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. -
Immunocytochemistry/Immunofluorescence analysis of A-673 (human muscle Ewing's Sarcoma cell line) cells labelling alpha smooth muscle Actin with purified ab124964 at 1/300. Cells were fixed with 4% paraformaldehyde. An Alexa Fluor® 555-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain.
Control: primary antibody (1/300) and secondary antibody, ab150113, an Alexa Fluor® 488-conjugated goat anti-mouse IgG (1/500).
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Different batches of ab124964 were tested on HEK-293 (Human embryonic kidney epithelial cell) lysate at 2.1 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 42 kDa.
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All lanes : Anti-alpha smooth muscle Actin antibody [EPR5368] (ab124964) at 1/10000 dilution (purified)
Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) cell lysate
Lane 2 : HEK-293 (human epithelial cell line from embryonic kidney) cell lysate
Lane 3 : U937 (human histiocytic lymphoma cell line) cell lysate
Lane 4 : A431 (human epidermoid carcinoma cell line) cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 42 kDa
Observed band size: 42 kDaBlocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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All lanes : Anti-alpha smooth muscle Actin antibody [EPR5368] (ab124964) at 1/50000 dilution (purified)
Lane 1 : Mouse brain tissue lysate
Lane 2 : Rat brain tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 42 kDa
Observed band size: 42 kDaBlocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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All lanes : Anti-alpha smooth muscle Actin antibody [EPR5368] (ab124964) at 1/10000 dilution
Lane 1 : Human lung lysate
Lane 2 : A431 (human epidermoid carcinoma cell line) cell lysate
Lane 3 : A549 (human lung carcinoma cell line) cell lysate
Lane 4 : NIH/3T3 (mouse embyro fibroblast cell line) cell lysate
Lane 5 : SV40-LT cell lysate
Lane 6 : C2C12 (mouse myoblast cell line) cell lysate
Lysates/proteins at 20 µg per lane.
Developed using the ECL technique.
Predicted band size: 42 kDa
Observed band size: 42 kDa
Exposure time: 10 secondsThis blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% milk before being incubated with ab124964 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406
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All lanes : Anti-alpha smooth muscle Actin antibody [EPR5368] (ab124964) at 1/1000 dilution (unpurified)
Lane 1 : Human heart tissue lysate - total protein (ab29431)
Lane 2 : Heart (Mouse) Tissue Lysate
Lane 3 : Heart (Rat) Tissue Lysate
Lane 4 : Human skeletal muscle tissue lysate - total protein (ab29330)
Lane 5 : HEK-293 (human epithelial cell line from embryonic kidney) Whole Cell Lysate
Lane 6 : A431 (human epidermoid carcinoma cell line) Whole Cell Lysate
Lane 7 : SV40LT-SMC (rat aorta smooth muscle) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 42 kDa
Additional bands at: 42 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 30 seconds -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human ovary tissue labelling alpha smooth muscle Actin with unpurified ab124964 at 1/1000 dilution.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human heart tissue labelling alpha smooth muscle actin with unpurified ab124964 at 1/1000 dilution. Note positive staining on smooth muscle cells but negative on striated muscle cells.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of normal human liver vessels tissue labelling alpha smooth muscle Actin with unpurified ab124964.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of normal human colon smooth muscle tissue labelling alpha smooth muscle Actin with unpurified ab124964.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostatic carcinoma smooth muscles tissue labelling alpha smooth muscle Actin with unpurified ab124964.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis showing vascular smooth muscle staining in skeletal muscle tissue using alpha smooth muscle Actin with unpurified ab124964. Note positive staining on smooth muscle cells but negative on striated muscle cells.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human stomach carcinoma smooth muscles tissue labelling alpha smooth muscle Actin with unpurified ab124964.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of normal human tonsil vessels tissue labelling alpha smooth muscle Actin with unpurified ab124964.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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ab124964 staining alpha smooth muscle Actin in Mouse Uterus tissue sections by Immunohistochemistry (Formalin/PFA perfusion fixed frozen sections). Tissue samples were fixed by perfusion with formaldehyde, blocked with PB ab64226 for 10 minutes at Room temperature and antigen retrieval was by heat mediation in citrate buffer. The sample was incubated with primary antibody (1/2000) for 30 minutes. A HRP-conjugated Goat anti-rabbit polyclonal (undiluted) was used as the secondary antibody.
プロトコール
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (289)
ab124964 は 289 報の論文で使用されています。
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- Chen K et al. Adipose-targeted triiodothyronine therapy counteracts obesity-related metabolic complications and atherosclerosis with negligible side effects. Nat Commun 13:7838 (2022). PubMed: 36539421