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alpha smooth muscle Actin Recombinant Rabbit Monoclonal Antibody [SY02-64] - BSA and Azide free

Usd: 649

Specification

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Catalog# HA750126

alpha smooth muscle Actin Recombinant Rabbit Monoclonal Antibody [SY02-64] - BSA and Azide free

Application

  • WB

  • IF-Cell

  • IF-Tissue

  • IHC-P

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

With BSA and Azide
Conjugation

  • unconjugated

Overview

Product Name

alpha smooth muscle Actin Recombinant Rabbit Monoclonal Antibody [SY02-64] - BSA and Azide free

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within N-terminal human alpha smooth muscle Actin.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IF-Tissue, IHC-P, FC

Molecular Weight

Predicted band size: 42 kDa

Positive Control

Saos-2 (Human osteosarcoma cells) cell lysate, A431 (Human epidermoid carcinoma skin squamous cells) cell lysate, NIH/3T3 (Mouse fibroblasts) cell lysate, C2C12 (Mouse myoblasts) cell lysate, Neuro-2a (Mouse brain neuroblastoma cells) cell lysate, Mouse skin tissue lysate, Rat skin tissue lysate.

Conjugation

unconjugated

Clone Number

SY02-64

Product Features

Form

Liquid

Concentration

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000-1:5,000

  • IF-Cell

  • 1:2,000-1:5,000

  • IF-Tissue

  • 1:50-1:200

  • IHC-P

  • 1:1,000-1:5,000

  • FC

  • 1:1,000-1:2,000

Target

Function

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.

Background References

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).

Sequence Similarity

Belongs to the actin family.

Post-translational Modification

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.

Subcellular Location

Cytoplasm.

UNIPROT

SwissProt: P62736 Human

SwissProt: P62737 Mouse

SwissProt: P62738 Rat

Synonyms

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

Expand

Images

  • Application: IF-tissue<br /><br />Species: Human<br /><br />Site: Kidney <br /><br />Sample: Paraffin-embedded section<br /><br />Antibody concentration: 1/50

    Application: IF-tissue

    Species: Human

    Site: Kidney

    Sample: Paraffin-embedded section

    Antibody concentration: 1/50

  • Application: IF-tissue<br /><br />Species: Human<br /><br />Site: Stomach cancer<br /><br />Sample: Paraffin-embedded section<br /><br />Antibody concentration: 1/50 (alpha smooth muscle Actin, <a href="/products/HA750126" style="font-weight: bold;text-decoration: underline;">HA750126</a>, red) and 1/100 (ALDH2, <a href="/products/M1509-1" style="font-weight: bold;text-decoration: underline;">M1509-1</a>, green)

    Application: IF-tissue

    Species: Human

    Site: Stomach cancer

    Sample: Paraffin-embedded section

    Antibody concentration: 1/50 (alpha smooth muscle Actin, HA750126, red) and 1/100 (ALDH2, M1509-1, green)

  • Application: IF-tissue<br /><br />Species: Mouse<br /><br />Site: Liver<br /><br />Sample: Paraffin-embedded section<br /><br />Antibody concentration: 1/50

    Application: IF-tissue

    Species: Mouse

    Site: Liver

    Sample: Paraffin-embedded section

    Antibody concentration: 1/50

  • Western blot analysis of alpha smooth muscle Actin on different lysates with Rabbit anti-alpha smooth muscle Actin antibody (<a href="/products/HA750126" style="font-weight: bold;text-decoration: underline;">HA750126</a>) at 1/2,000 dilution.<br /><br />Lane 1: Saos-2 (Human osteosarcoma cells) cell lysate<br />Lane 2: A431 (Human epidermoid carcinoma skin squamous cells) cell lysate<br />Lane 3: NIH/3T3 (Mouse fibroblasts) cell lysate<br />Lane 4: C2C12 (Mouse myoblasts) cell lysate<br />Lane 5: Neuro-2a (Mouse brain neuroblastoma cells) cell lysate<br />Lane 6: Mouse skin tissue lysate<br />Lane 7: Rat skin tissue lysate<br /><br />Lysates/proteins at 15 µg/Lane.<br />Exposure time: 2 seconds; ECL: K1801<br /><br />Blocking: 5% NFDM/TBST, 1 hour at room temperature<br />Primary antibody: <a href="/products/HA750126" style="font-weight: bold;text-decoration: underline;">HA750126</a>, 1/2,000 in primary antibody dilution buffer (<a href="/products/K1803" style="font-weight: bold;text-decoration: underline;">K1803</a>), overnight at 4 ℃<br />Secondary antibody: Goat anti-Rabbit IgG-HRP (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature<br /><br />Predicted band size: 42 kDa<br />Observed band size: 42 kDa

    Western blot analysis of alpha smooth muscle Actin on different lysates with Rabbit anti-alpha smooth muscle Actin antibody (HA750126) at 1/2,000 dilution.

    Lane 1: Saos-2 (Human osteosarcoma cells) cell lysate
    Lane 2: A431 (Human epidermoid carcinoma skin squamous cells) cell lysate
    Lane 3: NIH/3T3 (Mouse fibroblasts) cell lysate
    Lane 4: C2C12 (Mouse myoblasts) cell lysate
    Lane 5: Neuro-2a (Mouse brain neuroblastoma cells) cell lysate
    Lane 6: Mouse skin tissue lysate
    Lane 7: Rat skin tissue lysate

    Lysates/proteins at 15 µg/Lane.
    Exposure time: 2 seconds; ECL: K1801

    Blocking: 5% NFDM/TBST, 1 hour at room temperature
    Primary antibody: HA750126, 1/2,000 in primary antibody dilution buffer (K1803), overnight at 4 ℃
    Secondary antibody: Goat anti-Rabbit IgG-HRP (HA1001), 1/50,000 in 5% NFDM/TBST, 1 hour at room temperature

    Predicted band size: 42 kDa
    Observed band size: 42 kDa

  • Application: Immunocytochemistry (IF-cell)<br /><br />Species: Human<br />Sample: Hep G2 (Human liver cancer cells)<br /><br />Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.<br />Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.<br /><br />Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.<br />Antibody dilution buffer: 1% BSA in PBST.<br />Primary antibody: <a href="/products/HA750126" style="font-weight: bold;text-decoration: underline;">HA750126</a>, 1/2,000, overnight at 4℃.<br />Secondary antibody: Goat Anti-Rabbit IgG (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>), 45 minutes at room temperature.<br /><br />Counterstain: Beta tubulin (<a href="/products/HA601187" style="font-weight: bold;text-decoration: underline;">HA601187</a>, red), 1/100, overnight at 4℃. The Nuclear counterstain was DAPI (Blue).

    Application: Immunocytochemistry (IF-cell)

    Species: Human
    Sample: Hep G2 (Human liver cancer cells)

    Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
    Permeabilization: 0.1% Triton X-100, 15 minutes at room temperature.

    Blocking: 1% BSA + 10% normal goat serum, 1 hour at room temperature.
    Antibody dilution buffer: 1% BSA in PBST.
    Primary antibody: HA750126, 1/2,000, overnight at 4℃.
    Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 45 minutes at room temperature.

    Counterstain: Beta tubulin (HA601187, red), 1/100, overnight at 4℃. The Nuclear counterstain was DAPI (Blue).

  • Application: Immunohistochemistry (IHC-P)<br /><br />Species: Human<br />Tissue: Breast carcinoma<br />Sample: Paraffin-embedded section<br /><br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.<br /><br />Wash buffer: 1× TBST<br />Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.<br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.<br />Primary antibody: <a href="/products/HA750126" style="font-weight: bold;text-decoration: underline;">HA750126</a>, 1/1,000, 1 hour at room temperature.<br />Secondary antibody: <a href="/products/HA1119" style="font-weight: bold;text-decoration: underline;">HA1119</a>, 20 minutes at room temperature.

    Application: Immunohistochemistry (IHC-P)

    Species: Human
    Tissue: Breast carcinoma
    Sample: Paraffin-embedded section

    Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.

    Wash buffer: 1× TBST
    Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
    Primary antibody: HA750126, 1/1,000, 1 hour at room temperature.
    Secondary antibody: HA1119, 20 minutes at room temperature.

  • Application: Immunohistochemistry (IHC-P)<br /><br />Species: Mouse<br />Tissue: Small intestine<br />Sample: Paraffin-embedded section<br /><br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.<br /><br />Wash buffer: 1× TBST<br />Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.<br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.<br />Primary antibody: <a href="/products/HA750126" style="font-weight: bold;text-decoration: underline;">HA750126</a>, 1/1,000, 1 hour at room temperature.<br />Secondary antibody: <a href="/products/HA1119" style="font-weight: bold;text-decoration: underline;">HA1119</a>, 20 minutes at room temperature.

    Application: Immunohistochemistry (IHC-P)

    Species: Mouse
    Tissue: Small intestine
    Sample: Paraffin-embedded section

    Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.

    Wash buffer: 1× TBST
    Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
    Primary antibody: HA750126, 1/1,000, 1 hour at room temperature.
    Secondary antibody: HA1119, 20 minutes at room temperature.

  • Application: Immunohistochemistry (IHC-P)<br /><br />Species: Rat<br />Tissue: Small intestine<br />Sample: Paraffin-embedded section<br /><br />Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.<br /><br />Wash buffer: 1× TBST<br />Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.<br />Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.<br />Primary antibody: <a href="/products/HA750126" style="font-weight: bold;text-decoration: underline;">HA750126</a>, 1/1,000, 1 hour at room temperature.<br />Secondary antibody: <a href="/products/HA1119" style="font-weight: bold;text-decoration: underline;">HA1119</a>, 20 minutes at room temperature.

    Application: Immunohistochemistry (IHC-P)

    Species: Rat
    Tissue: Small intestine
    Sample: Paraffin-embedded section

    Antigen retrieval: Heat-mediated, Tris-EDTA buffer (pH 9.0), 20 minutes at 95℃.

    Wash buffer: 1× TBST
    Endogenous peroxidase blocking: 3% H₂O₂, 10 minutes at room temperature.
    Blocking: 1% BSA + 10% normal goat serum, 10 minutes at room temperature.
    Primary antibody: HA750126, 1/1,000, 1 hour at room temperature.
    Secondary antibody: HA1119, 20 minutes at room temperature.

  • Application: Flow Cytometry (Intra)<br /><br />Species: Human<br />Sample: Hep G2 (Human liver cancer cells)<br /><br />Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.<br />Permeabilization: 1x Permeabilization Buffer (eBioscience), 15 minutes at room temperature.<br /><br />Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature.<br />Antibody dilution buffer: 1x PBS.<br />Primary antibody: <a href="/products/HA750126" style="font-weight: bold;text-decoration: underline;">HA750126</a> (1/1,000) (Red) compared with Rabbit IgG Isotype Control (<a href="/products/HA722127" style="font-weight: bold;text-decoration: underline;">HA722127</a>, Green), 15 minutes at room temperature.<br />Secondary antibody: Goat Anti-Rabbit IgG (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>), 15 minutes at room temperature.

    Application: Flow Cytometry (Intra)

    Species: Human
    Sample: Hep G2 (Human liver cancer cells)

    Fixation: 4% Paraformaldehyde, 15 minutes at room temperature.
    Permeabilization: 1x Permeabilization Buffer (eBioscience), 15 minutes at room temperature.

    Blocking: 1% BSA + 10% normal goat serum, 15 minutes at room temperature.
    Antibody dilution buffer: 1x PBS.
    Primary antibody: HA750126 (1/1,000) (Red) compared with Rabbit IgG Isotype Control (HA722127, Green), 15 minutes at room temperature.
    Secondary antibody: Goat Anti-Rabbit IgG (iFluor™ 488, HA1121), 15 minutes at room temperature.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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