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Actin Recombinant Rabbit Monoclonal Antibody [JJ09-29]

Usd: 385

Specification

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Catalog# ET1701-80

Actin Recombinant Rabbit Monoclonal Antibody [JJ09-29]

Application

  • WB

  • IF-Cell

  • IF-Tissue

  • IHC-P

Reactivity

  • Human

  • Mouse

  • Rat

  • Zebrafish

BSA and Azide free
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

Actin Recombinant Rabbit Monoclonal Antibody [JJ09-29]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide within Human Actin aa 45-80 / 377.

Species Reactivity

Human, Mouse, Rat, Zebrafish

Validated Applications

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

Molecular Weight

Predicted band size: 42 kDa

Positive Control

HeLa cell lysate, A431 cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, mouse brain tissue lysate, mouse colon tissue lysate, rat brain tissue lysate, rat colon tissue lysate, human colon carcinoma tissue, hybrid fish (crucian-carp) brain tissue lysate, hybrid fish (crucian-carp) kidney tissue lysate, NIH/3T3, mouse cardiac muscle tissue, mouse smooth muscle tissue.

Conjugation

unconjugated

Clone Number

JJ09-29

RRID

AB_2943481

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:10,000-1:50,000

  • IF-Cell

  • 1:100-1:500

  • IF-Tissue

  • 1:1,000

  • IHC-P

  • 1:5,000

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. Moilanen AM et al. WDR12, a Member of Nucleolar PeBoW-Complex, Is Up-Regulated in Failing Hearts and Causes Deterioration of Cardiac Function. PLoS One 10:e0124907 (2015).

2. Rafatian N et al. Cardiac macrophages and apoptosis after myocardial infarction: effects of central MR blockade. Am J Physiol Regul Integr Comp Physiol 307:R879-87 (2014).

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, Cytoskeleton.

UNIPROT

SwissProt: P68133 Human

SwissProt: P68134 Mouse

SwissProt: P68136 Rat

Entrez Gene: 407658 Zebrafish

Synonyms

a actin antibody

ACTA antibody

ACTA1 antibody

Actin alpha skeletal muscle antibody

Actin antibody

actin, alpha 1, skeletal muscle 1 antibody

actin, alpha 1, skeletal muscle antibody

Actin, alpha skeletal muscle antibody

actina antibody

actine antibody

Expand

Images

  • Western blot analysis of Actin on different lysates with Rabbit anti-Actin antibody (<a href="/products/ET1701-80" style="font-weight: bold;text-decoration: underline;">ET1701-80</a>) at 1/10,000 dilution.<br /><br />Lane 1: HeLa cell lysate (15 µg/Lane)<br />Lane 2: A431 cell lysate (15 µg/Lane)<br />Lane 3: NIH/3T3 cell lysate (15 µg/Lane)<br />Lane 4: PC-12 cell lysate (15 µg/Lane)<br />Lane 5: Mouse brain tissue lysate (20 µg/Lane)<br />Lane 6: Mouse colon tissue lysate (20 µg/Lane)<br />Lane 7: Rat brain tissue lysate (20 µg/Lane)<br />Lane 8: Rat colon tissue lysate (20 µg/Lane)<br /><br />Predicted band size: 42 kDa<br />Observed band size: 42 kDa<br /><br />Exposure time: 24 seconds;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1701-80" style="font-weight: bold;text-decoration: underline;">ET1701-80</a>) at 1/10,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/100,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of Actin on different lysates with Rabbit anti-Actin antibody (ET1701-80) at 1/10,000 dilution.

    Lane 1: HeLa cell lysate (15 µg/Lane)
    Lane 2: A431 cell lysate (15 µg/Lane)
    Lane 3: NIH/3T3 cell lysate (15 µg/Lane)
    Lane 4: PC-12 cell lysate (15 µg/Lane)
    Lane 5: Mouse brain tissue lysate (20 µg/Lane)
    Lane 6: Mouse colon tissue lysate (20 µg/Lane)
    Lane 7: Rat brain tissue lysate (20 µg/Lane)
    Lane 8: Rat colon tissue lysate (20 µg/Lane)

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

    Exposure time: 24 seconds;

    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 (ET1701-80) at 1/10,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/100,000 dilution was used for 1 hour at room temperature.

  • Application: IF-tissue<br /><br />Species: Human<br /><br />Site: Colon carcinoma<br /><br />Sample: Paraffin-embedded section<br /><br />Antibody concentration: 1/100

    Application: IF-tissue

    Species: Human

    Site: Colon carcinoma

    Sample: Paraffin-embedded section

    Antibody concentration: 1/100

  • Western blot analysis of Actin on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (<a href="/products/ET1701-80" style="font-weight: bold;text-decoration: underline;">ET1701-80</a>, 1/10,000) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.<br />Positive control:<br />Lane 1: Hybrid fish (crucian-carp) brain tissue lysate<br />Lane 2: Hybrid fish (crucian-carp) kidney tissue lysate

    Western blot analysis of Actin on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1701-80, 1/10,000) was 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.
    Positive control:
    Lane 1: Hybrid fish (crucian-carp) brain tissue lysate
    Lane 2: Hybrid fish (crucian-carp) kidney tissue lysate

  • Immunocytochemistry analysis of NIH/3T3 cells labeling Actin with Rabbit anti-Actin antibody (<a href="/products/ET1701-80" style="font-weight: bold;text-decoration: underline;">ET1701-80</a>) at 1/100 dilution.<br /><br />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-Actin antibody (<a href="/products/ET1701-80" style="font-weight: bold;text-decoration: underline;">ET1701-80</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) 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.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of NIH/3T3 cells labeling Actin with Rabbit anti-Actin antibody (ET1701-80) at 1/100 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-Actin antibody (ET1701-80) at 1/100 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.

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

Citation

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