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肌動蛋白α/α-SMA/α Actin單克隆抗體

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產品編號bsm-33187M
英文名稱Mouse Anti-alpha smooth muscle Actin antibody
中文名稱肌動蛋白α/α-SMA/α Actin單克隆抗體
別    名alpha sarcomeric Actin; alpha smooth muscle Actin; Actin alpha; ASMA; ASM-A; alpha-SMA; alpha SMA; AAT6; ACTA2; Actin alpha 2 smooth muscle aorta; Actin aortic smooth muscle; ACTSA; ACTVS; Alpha 2 actin; Alpha-actin 2; Cell growth inhibiting gene 46 protein; Growth inhibiting gene 46; ACTA_HUMAN; Actin alpha 2 smooth muscle aorta; Actin aortic smooth muscle; Actin, aortic smooth muscle; Alpha 2 actin; Alpha actin 2; Alpha cardiac actin; Alpha-actin 2; Alpha-actin-2; Cell growth inhibiting gene 46 protein; Cell growth-inhibiting gene 46 protein; Growth inhibiting gene 46; MYMY5  α-Smooth Muscle Actin; α Smooth Muscle Actin;
Specific References  (11)     |     bsm-33187M has been referenced in 11 publications.
[IF=10.435] Mei, Jiawei. et al. An injectable photo-cross-linking silk hydrogel system augments diabetic wound healing in orthopaedic surgery through spatiotemporal immunomodulation. J NANOBIOTECHNOL. 2022 Dec;20(1):1-22  IHC ;  Mouse.  
[IF=6.107] Lianglong Chen. et al. Multifunctional sponge scaffold loaded with concentrated growth factors for promoting wound healing. ISCIENCE. 2023 Jan;26:105835  IHC ;  Rat.  
[IF=5.696] Jiang-Tao Fan. et al. Exosomal lncRNA NEAT1 from cancer-associated fibroblasts facilitates endometrial cancer progression via miR-26a/b-5p-mediated STAT3/YKL-40 signaling pathway. Neoplasia. 2021 Jul;23:692  IF ;  Human.  
[IF=4.967] Weijie Jiao. et al. Construction and Evaluation of Small-Diameter Bioartificial Arteries Based on a Combined-Mold Technology. POLYMERS-BASEL. 2022 Jan;14(15):3089  IF ;  Rat.  
[IF=4.101] Zengxian Sun. et al. Metformin inhibits pulmonary artery smooth muscle cell proliferation by upregulating p21 via NONRATT015587.2. Int J Mol Med. 2022 Apr;49(4):1-13  FC ;  Rat.  
[IF=4.068] Ming Li. et al. ADAMTS12, a novel prognostic predictor, promotes cell proliferation, migration and invasion in head and neck squamous cell carcinoma. ORAL DIS. 2022 Oct;:  IF ;  Human.  
[IF=3.61] Lu X. et al. Anti-renal fibrosis effect of asperulosidic acid via TGF-β1/smad2/smad3 and NF-κB signaling pathways in a rat model of unilateral ureteral obstruction  IHC ;  human.  
[IF=3.067] Liyan Shi. et al. Cranberry (Vacinium macrocarpon) phytochemicals inhibit hepatic stellate cell activation and liver fibrosis. Food Biosci. 2021 Aug;42:101176  IF ;  Rat.  
[IF=3.046] Tan HX et al. TGFβ1 is essential for MSCs-CAFs differentiation and promotes HCT116 cells migration and invasion via JAK/STAT3 signaling. Onco Targets Ther. 2019 Jul 5;12:5323-5334.  FCM ;  Human.  
[IF=2.751] Ziqian Liu. et al. Eplerenone ameliorates lung fibrosis in unilateral ureteral obstruction rats by inhibiting lymphangiogenesis. EXP THER MED. 2022 Oct;24(4):1-12  IHC ;  Rat.  
[IF=1.56] Jin, Yingli, et al. "Urinary kidney injury molecule?1 as an early diagnostic biomarker of obstructive acute kidney injury and development of a rapid detection method." Molecular Medicine Reports.  IHC-P ;  Rat.  
研究領域細胞生物  發育生物學  細胞骨架  
抗體來源Mouse
克隆類型Monoclonal
克 隆 號3F9
交叉反應Rat,Mouse,Human
產品應用WB=1:500-5000, IHC-P=1:200-800, IHC-F=1:100-500, IF=1:100-500, Flow-Cyt=1ug/Test
not yet tested in other applications.
optimal dilutions/concentrations should be determined by the end user.
理論分子量42kDa
細胞定位細胞漿 
性    狀Liquid
濃    度1mg/ml
免 疫 原KLH conjugated synthetic peptide derived from human Actin alpha 
亞    型IgG
純化方法affinity purified by Protein G
緩 沖 液0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol.
保存條件Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.
注意事項This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.
PubMedPubMed
產品介紹The product encoded by this gene belongs to the actin family of proteins, which are highly conserved proteins that play a role in cell motility, structure and integrity. Alpha, beta and gamma actin isoforms have been identified, with alpha actins being a major constituent of the contractile apparatus, while beta and gamma actins are involved in the regulation of cell motility. This actin is an alpha actin that is found in skeletal muscle. Mutations in this gene cause nemaline myopathy type 3, congenital myopathy with excess of thin myofilaments, congenital myopathy with cores, and congenital myopathy with fiber-type disproportion, diseases that lead to muscle fiber defects. [provided by RefSeq, Jul 2008]

Function:
Actins are highly conserved proteins that are involved in various types of cell motility and are ubiquitously expressed in all eukaryotic cells.

Subunit:
Polymerization of globular actin (G-actin) leads to a structural filament (F-actin) in the form of a two-stranded helix. Each actin can bind to 4 others.

Subcellular Location:
Cytoplasm, cytoskeleton.

Post-translational modifications:
Oxidation of Met-46 by MICALs (MICAL1, MICAL2 or MICAL3) to form methionine sulfoxide promotes actin filament depolymerization. Methionine sulfoxide is produced stereospecifically, but it is not known whether the (S)-S-oxide or the (R)-S-oxide is produced (By similarity).

DISEASE:
Note=ACTA2 mutations predispose patients to a variety of diffuse and diverse vascular diseases, premature onset coronary artery disease (CAD), premature ischemic strokes and Moyamoya disease.
Defects in ACTA2 are the cause of familial aortic aneurysm 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.
Defects in ACTA2 are the cause of Moyamoya disease type 5 (MYMY5) [MIM:614042]. Moyamoya disease is a progressive cerebral angiopathy characterized by bilateral intracranial carotid artery stenosis and telangiectatic vessels in the region of the basal ganglia. The abnormal vessels resemble a 'puff of smoke' (moyamoya) on cerebral angiogram. Affected individuals can develop transient ischemic attacks and/or cerebral infarction, and rupture of the collateral vessels can cause intracranial hemorrhage. Hemiplegia of sudden onset and epileptic seizures constitute the prevailing presentation in childhood, while subarachnoid bleeding occurs more frequently in adults.
Defects in ACTA2 are the cause of multisystemic smooth muscle dysfunction syndrome (MSMDYS) [MIM:613834]. MSMDYS is a syndrome characterized by dysfunction of smooth muscle cells throughout the body, leading to aortic and cerebrovascular disease, fixed dilated pupils, hypotonic bladder, malrotation, and hypoperistalsis of the gut and pulmonary hypertension.

Similarity:
Belongs to the actin family.

SWISS:
P62736

Gene ID:
59

Database links:
Entrez Gene: 101021287 Baboon

Entrez Gene: 515610 Cow

Entrez Gene: 59 Human

Entrez Gene: 11475 Mouse

Entrez Gene: 733615 Pig

Entrez Gene: 100009271 Rabbit

Entrez Gene: 81633 Rat

Omim: 102620 Human

SwissProt: P62739 Cow

SwissProt: P62736 Human

SwissProt: P62737 Mouse

SwissProt: P62740 Rabbit

SwissProt: P62738 Rat

Unigene: 500483 Human

Unigene: 213025 Mouse

Unigene: 195319 Rat

Unigene: 3114 Rat



結構蛋白(Structural Proteins)
Actin α/α-Actin 是一種具有收縮能力的微絲蛋白,a-SMA廣泛分布于幾乎所有的肌型細胞中。Actin-α蛋白主要用于檢測骨骼肌、平滑肌、血管平滑肌、心肌和肌原性腫瘤 包括:平滑肌瘤、平滑肌肉瘤、橫紋肌肉瘤以及肌上細胞和肌上皮瘤。Actin(肌動蛋白)是在所有真核細胞中都表達的高度保守的蛋白質。它們沿微管組成了細胞骨架的主要成分。肌動蛋白至少表達為6種異構形式。它在心臟、骨骼橫紋肌組織和某些平滑肌組織中表達,調節其收縮功能。有報導說肌動蛋白在乳房瘤中是高度磷酸化的。肌動蛋白的功能失調也會導致某種類型的心臟病。平滑肌α肌動蛋白使人更感興趣,因為編碼它的基因是相對局限于在血管平滑肌細胞中表達的少數幾個基因之一。肌動蛋白是標記平滑肌和肌上皮細胞腫瘤的有效工具。
產品圖片
Sample:
Lane 1: Large intestine (Mouse) Lysate at 40 ug
Lane 2: Stomach (Mouse) Lysate at 40 ug
Lane 3: Uterus (Mouse) Lysate at 40 ug
Lane 4: Kidney (Mouse) Lysate at 40 ug
Lane 5: Small intestine (Mouse) Lysate at 40 ug
Lane 6: NIH/3T3 (Mouse) Cell Lysate at 30 ug
Lane 7: Stomach (Rat) Lysate at 40 ug
Lane 8: Uterus (Rat) Lysate at 40 ug
Lane 9: A549 (Human) Cell Lysate at 30 ug
Lane 10: Hela (Human) Cell Lysate at 30 ug
Lane 11: A431 (Human) Cell Lysate at 30 ug
Lane 12: HepG2 (Human) Cell Lysate at 30 ug
Primary: Anti-alpha smooth muscle Actin (bsm-33187M) at 1/1000 dilution
Secondary: IRDye800CW Goat Anti-Mouse IgG at 1/20000 dilution
Predicted band size: 42 kD
Observed band size: 42 kD
Paraformaldehyde-fixed, paraffin embedded (Human uterus); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (bsm-33187M) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human cervical carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (cell supernatant of bsm-33187M 7F9) at overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat small intestine); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (cell supernatant of bsm-33187M 7F9) at overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat uterus); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (bsm-33187M) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (mouse uterus); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (bsm-33187M) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (mouse stomach); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (bsm-33187M) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat stomach); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (bsm-33187M) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human colon carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (cell supernatant of bsm-33187M 7F9) at overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat stomach); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (cell supernatant of bsm-33187M 7F9) at overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat colon); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (cell supernatant of bsm-33187M 7F9) at overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (mouse stomach); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (cell supernatant of bsm-33187M 7F9) at overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (Mouse small intestine); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (cell supernatant of bsm-33187M 7F9) at overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (mouse colon); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (cell supernatant of bsm-33187M 7F9) at overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (Human smooth muscle); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (cell supernatant of bsm-33187M 7F9) at overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat uterus); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (cell supernatant of bsm-33187M 7F9) at overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human colon carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (bsm-33187M) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat small intestine); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (bsm-33187M) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat colon); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (bsm-33187M) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (Mouse small intestine); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (bsm-33187M) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (mouse colon); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (bsm-33187M) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat stomach); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (ascites of bsm-33187M 3F9) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Mouse) (sp-0024) instructions and DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human gastric); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (ascites of bsm-33187M 3F9) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Mouse) (sp-0024) instructions and DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human colon carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (ascites of bsm-33187M 3F9) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Mouse) (sp-0024) instructions and DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat kidney); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (ascites of bsm-33187M 3F9) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Mouse) (sp-0024) instructions and DAB staining.
Paraformaldehyde-fixed, paraffin embedded (human cervical carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (ascites of bsm-33187M 3F9) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Mouse) (sp-0024) instructions and DAB staining.
Paraformaldehyde-fixed, paraffin embedded (rat colon); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (ascites of bsm-33187M 3F9) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Mouse) (sp-0024) instructions and DAB staining.
Paraformaldehyde-fixed, paraffin embedded (Mouse small intestine); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (ascites of bsm-33187M 3F9) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Mouse) (sp-0024) instructions and DAB staining.
Paraformaldehyde-fixed, paraffin embedded (Mouse colon); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (ascites of bsm-33187M 3F9) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Mouse) (sp-0024) instructions and DAB staining.
Paraformaldehyde-fixed, paraffin embedded (mouse uterus); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (cell supernatant of bsm-33187M 7F9) at overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining.
Paraformaldehyde-fixed, paraffin embedded (Rat stomach); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (alpha smooth muscle Actin) Monoclonal Antibody, Unconjugated (bsm-33187M) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Mouse) (sp-0024) instructionsand DAB staining.
Blank control:NIH/3T3.
Primary Antibody (green line): Mouse Anti-alpha smooth muscle Actin antibody (bsm-33187M)
Dilution: 1ug/Test;
Secondary Antibody : Goat anti-mouse IgG-FITC
Dilution: 0.5ug/Test.
Protocol
The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at -20℃.The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.
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