alpha smooth muscle Actin Recombinant Rabbit Monoclonal Antibody [SY02-64] - BSA and Azide free
Catalog# HA750126
alpha smooth muscle Actin Recombinant Rabbit Monoclonal Antibody [SY02-64] - BSA and Azide free
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WB
-
IF-Cell
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IF-Tissue
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IHC-P
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FC
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Human
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Mouse
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Rat
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ET1607-53
含抗保成分
-
unconjugated
概述
产品名称
alpha smooth muscle Actin Recombinant Rabbit Monoclonal Antibody [SY02-64] - BSA and Azide free
抗体类型
Recombinant Rabbit monoclonal Antibody
免疫原
Synthetic peptide within N-terminal human alpha smooth muscle Actin.
种属反应性
Human, Mouse, Rat
验证应用
WB, IF-Cell, IF-Tissue, IHC-P, FC
分子量
Predicted band size: 42 kDa
阳性对照
Saos-2 cell lysate, A431 cell lysate, NIH/3T3 cell lysate, C2C12 cell lysate, Neuro-2a cell lysate, Mouse skin tissue lysate, Rat skin tissue lysate, human breast carcinoma tissue, NIH/3T3, HUVEC, human kidney tissue, human small intestine tissue, mouse small intestine tissue, rat small intestine tissue, HepG2.
偶联
unconjugated
克隆号
SY02-64
产品特性
形态
Liquid
存放说明
Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
存储缓冲液
PBS (pH7.4).
亚型
IgG
纯化方式
Protein A affinity purified.
应用稀释度
-
WB
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1:20,000-1:50,000
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IF-Cell
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1:2,000-1:5,000
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IF-Tissue
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1:1,000-1:2,000
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IHC-P
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1:50,000-1:100,000
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FC
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1:1,000-1:2,000
靶点
功能
All eukaryotic cells express Actin, which often constitutes as much as 50% of total cellular protein. Actin filaments can form both stable and labile structures and are crucial components of microvilli and the contractile apparatus of muscle cells. While lower eukaryotes, such as yeast, have only one Actin gene, higher eukaryotes have several isoforms encoded by a family of genes. At least six types of Actin are present in mammalian tissues and fall into three classes. α-Actin expression is limited to various types of muscle, whereas β-Actin and γ-Actin are the principle constituents of filaments in other tissues. Members of the small GTPase family regulate the organization of the Actin cytoskeleton. Rho controls the assembly of Actin stress fibers and focal adhesion. Rac regulates Actin filament accumulation at the plasma membrane. Cdc42 stimulates formation of filopodia.
背景文献
1. Izumi D et al. CXCL12/CXCR4 activation by cancer-associated fibroblasts promotes integrin 1 clustering and invasiveness in gastric cancer. Int J Cancer 138:1207-19 (2016).
2. Chung SI et al. Development of a transgenic mouse model of hepatocellular carcinoma with a liver fibrosis background. BMC Gastroenterol 16:13 (2016).
序列相似性
Belongs to the actin family.
翻译后修饰
Oxidation of Met-46 and Met-49 by MICALs (MICAL1, MICAL2 or MICAL3) to form methionine sulfoxide promotes actin filament depolymerization. MICAL1 and MICAL2 produce the (R)-S-oxide form. The (R)-S-oxide form is reverted by MSRB1 and MSRB2, which promotes actin repolymerization.; Monomethylation at Lys-86 (K84me1) regulates actin-myosin interaction and actomyosin-dependent processes. Demethylation by ALKBH4 is required for maintaining actomyosin dynamics supporting normal cleavage furrow ingression during cytokinesis and cell migration.; Methylated at His-75 by SETD3.; (Microbial infection) Monomeric actin is cross-linked by V.cholerae toxins RtxA and VgrG1 in case of infection: bacterial toxins mediate the cross-link between Lys-52 of one monomer and Glu-272 of another actin monomer, resulting in formation of highly toxic actin oligomers that cause cell rounding. The toxin can be highly efficient at very low concentrations by acting on formin homology family proteins: toxic actin oligomers bind with high affinity to formins and adversely affect both nucleation and elongation abilities of formins, causing their potent inhibition in both profilin-dependent and independent manners.
亚细胞定位
Cytoplasm.
别名
alpha SMA
a-SMA antibody
asma antibody
a actin antibody
AAT6 antibody
ACTA_HUMAN antibody
ACTA2 antibody
Actin alpha 2 smooth muscle aorta antibody
Actin aortic smooth muscle antibody
Actin, aortic smooth muscle antibody
展开图片
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Western blot analysis of alpha smooth muscle Actin on different lysates with Rabbit anti-alpha smooth muscle Actin antibody (HA750126) at 1/20,000 dilution and competitor's antibody at 1/5,000 dilution.
Lane 1: Saos-2 cell lysate (15 µg/Lane)
Lane 2: A431 cell lysate (15 µg/Lane)
Lane 3: NIH/3T3 cell lysate (15 µg/Lane)
Lane 4: C2C12 cell lysate (15 µg/Lane)
Lane 5: Neuro-2a cell lysate (15 µg/Lane)
Lane 6: Mouse skin tissue lysate (30 µg/Lane)
Lane 7: Rat skin tissue lysate (30 µg/Lane)
Predicted band size: 42 kDa
Observed band size: 42 kDa
Exposure time: 46 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA750126) at 1/20,000 dilution and competitor's antibody at 1/5,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
☑ Knockdown (KD)
Western blot analysis of alpha smooth muscle Actin on different lysates with Rabbit anti-alpha smooth muscle Actin antibody (HA750126) at 1/20,000 dilution.
Lane 1: Hela-si NT cell lysate (10 µg/Lane)
Lane 2: Hela-si alpha smooth muscle Actin cell lysate (10 µg/Lane)
Predicted band size: 42 kDa
Observed band size: 42 kDa
Exposure time: 5 seconds;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (ET1607-53, 1/20,000) and Loading control antibody (Rabbit anti-GAPDH, ET1601-4, 1/10,000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-rabbit IgG-HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue with Rabbit anti-alpha smooth muscle Actin antibody (HA750126) at 1/50,000 dilution and competitor's antibody at 1/20,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) (high pressure) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750126) at 1/50,000 dilution and competitor's antibody at 1/20,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Application: IF-tissue
Species: Human
Site: Kidney
Sample: Paraffin-embedded section
Antibody concentration: 1/1,000 -
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☑ Relative expression (RE)
Immunocytochemistry analysis of HepG2 (positive) and A431 (negative) labeling alpha smooth muscle Actin with Rabbit anti-alpha smooth muscle Actin antibody (HA750126) at 1/2,000 dilution.
Cells were fixed in 100% precooled methanol for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-alpha smooth muscle Actin antibody (HA750126) at 1/2,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. -
Immunocytochemistry analysis of NIH/3T3 cells labeling alpha smooth muscle Actin with Rabbit anti-alpha smooth muscle Actin antibody (HA750126) at 1/2,500 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-alpha smooth muscle Actin antibody (HA750126) at 1/2,500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunocytochemistry analysis of HUVEC cells labeling alpha smooth muscle Actin with Rabbit anti-alpha smooth muscle Actin antibody (HA750126) at 1/5,000 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-alpha smooth muscle Actin antibody (HA750126) at 1/5,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-alpha smooth muscle Actin antibody (HA750126) at 1/50,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750126) at 1/50,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded human small intestine tissue with Rabbit anti-alpha smooth muscle Actin antibody (HA750126) at 1/50,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750126) at 1/50,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded mouse small intestine tissue with Rabbit anti-alpha smooth muscle Actin antibody (HA750126) at 1/50,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750126) at 1/50,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded rat small intestine tissue with Rabbit anti-alpha smooth muscle Actin antibody (HA750126) at 1/50,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750126) at 1/50,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Application: IF-tissue
Species: Mouse
Site: Liver
Sample: Paraffin-embedded section
Antibody concentration: 1/1,000 -
Flow cytometric analysis of HepG2 cells labeling alpha smooth muscle Actin.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA750126, 1/2,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
请注意: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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