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beta Actin Mouse Monoclonal Antibody [A2-F6]

Usd: 350

Specification

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Catalog# EM21002

beta Actin Mouse Monoclonal Antibody [A2-F6]

Application

  • WB

  • IF-Cell

  • IHC-P

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

Conjugation

  • unconjugated

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

Overview

Product Name

beta Actin Mouse Monoclonal Antibody [A2-F6]

Antibody Type

Mouse Monoclonal Antibody

Immunogen

Synthetic peptide (KLH-coupled) within human Beta-actin aa 1-50.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IHC-P, FC

Molecular Weight

Predicted band size: 42 kDa

Positive Control

Hela cell lysate, THP-1 cell lysate, HepG2 cell lysate, MCF-7 cell lysate, PC-12 cell lysate, PMVEC cell lysate, RH-35 cell lysate, L6 cell lysate, NIH/3T3 cell lysate, L929 cell lysate, RAW264.7 cell lysate, B16F1 cell lysate, MEF cell lysate, Hela, A549, NIH/3T3, human colon carcinoma tissue, mouse prostate tissue, mouse kidney tissue.

Conjugation

unconjugated

Clone Number

A2-F6

RRID

AB_2819164

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*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG1

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:40,000-100,000

  • IF-Cell

  • 1:100-1:200

  • IHC-P

  • 1:1,000

  • FC

  • 1:1,000

Target

Function

Beta-actin (human gene and protein abbreviation ACTB/ACTB) is one of six different actin isoforms which have been identified in humans. This is one of the two nonmuscle cytoskeletal actins. Actins are highly conserved proteins[5][6] that are involved in cell motility, structure and integrity. Alpha actins are a major constituent of the contractile apparatus. Beta-actin has been shown to interact with SPTBN2. In addition, RNA-binding protein Sam68 was found to interact with the mRNA encoding β-actin, which regulates the synaptic formation of the dendritic spines with its cytoskeletal components. Beta-actin has been shown to activate eNOS, thereby increasing NO production. An eight-amino acid residue (326-333) in actin has been shown to mediate the interaction between actin and eNOS. Recurrent mutations in this gene have been associated to cases of diffuse large B-cell lymphoma. Beta actin is often used in Western blotting as a loading control, to normalize total protein amounts and check for eventual protein degradation in the samples. Its transcript is also commonly used as a housekeeping gene standard in qPCR. Its molecular weight is approximately 42 kDa.

Background References

1. Fujiki R et al. GlcNAcylation of a histone methyltransferase in retinoic-acid-induced granulopoiesis. Nature 459:455-459 (2009).

2. Fujiki R et al. Retraction: GlcNAcylation of a histone methyltransferase in retinoic-acid-induced granulopoiesis. Nature 505:574-574 (2014).

Sequence Similarity

Belongs to the actin family.

Post-translational Modification

ISGylated.; Oxidation of Met-44 and Met-47 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 promote actin repolymerization.; Monomethylation at Lys-84 (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-73 by SETD3. Methylation at His-73 is required for smooth muscle contraction of the laboring uterus during delivery (By similarity).; [Actin, cytoplasmic 1, N-terminally processed]: N-terminal acetylation by NAA80 affects actin filament depolymerization and elongation, including elongation driven by formins. In contrast, filament nucleation by the Arp2/3 complex is not affected.; (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-50 of one monomer and Glu-270 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: P60709 Human

SwissProt: P60710 Mouse

SwissProt: P60711 Rat

Synonyms

A26C1A antibody

A26C1B antibody

ACTB antibody

ACTB_HUMAN antibody

Actin beta antibody

Actin cytoplasmic 1 antibody

Actin, cytoplasmic 1, N-terminally processed antibody

Actx antibody

b actin antibody

Beta cytoskeletal actin antibody

Expand

Images

  • Western blot analysis of beta Actin on Hela cell lysates with Mouse anti-beta Actin antibody (<a href="/products/EM21002" style="font-weight: bold;text-decoration: underline;">EM21002</a>).<br /><br />Hela cell lysates at 10 µg/Lane.<br /><br />Predicted band size: 42 kDa<br />Observed band size: 42 kDa<br /><br />12% 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/EM21002" style="font-weight: bold;text-decoration: underline;">EM21002</a>) at serial dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1:100,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of beta Actin on Hela cell lysates with Mouse anti-beta Actin antibody (EM21002).

    Hela cell lysates at 10 µg/Lane.

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

    12% 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 (EM21002) at serial dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:100,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of beta Actin on different lysates with Mouse anti-beta Actin antibody (<a href="/products/EM21002" style="font-weight: bold;text-decoration: underline;">EM21002</a>) at 1/40,000 dilution.<br /><br />Cell lysates at 10 µg/Lane, tissue lysates at 20 µg/Lane.<br /><br />Predicted band size: 42 kDa<br />Observed band size: 42 kDa<br /><br />Exposure time: 1 minute;<br /><br />12% 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/EM21002" style="font-weight: bold;text-decoration: underline;">EM21002</a>) at 1/40,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1:100,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of beta Actin on different lysates with Mouse anti-beta Actin antibody (EM21002) at 1/40,000 dilution.

    Cell lysates at 10 µg/Lane, tissue lysates at 20 µg/Lane.

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

    Exposure time: 1 minute;

    12% 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 (EM21002) at 1/40,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:100,000 dilution was used for 1 hour at room temperature.

  • Western blot analysis of beta Actin on different lysates with Mouse anti-beta Actin antibody (<a href="/products/EM21002" style="font-weight: bold;text-decoration: underline;">EM21002</a>) at 1/40,000 dilution.<br /><br />Lane 1: THP-1 cell lysate<br />Lane 2: HepG2 cell lysate<br />Lane 3: PMVEC cell lysate<br />Lane 4: NIH/3T3 cell lysate<br />Lane 5: L929 cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 42 kDa<br />Observed band size: 42 kDa<br /><br />Exposure time: 2 minutes;<br /><br />12% 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/EM21002" style="font-weight: bold;text-decoration: underline;">EM21002</a>) at 1/40,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (<a href="/products/HA1006" style="font-weight: bold;text-decoration: underline;">HA1006</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    Western blot analysis of beta Actin on different lysates with Mouse anti-beta Actin antibody (EM21002) at 1/40,000 dilution.

    Lane 1: THP-1 cell lysate
    Lane 2: HepG2 cell lysate
    Lane 3: PMVEC cell lysate
    Lane 4: NIH/3T3 cell lysate
    Lane 5: L929 cell lysate

    Lysates/proteins at 10 µg/Lane.

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

    Exposure time: 2 minutes;

    12% 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 (EM21002) at 1/40,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

  • Immunocytochemistry analysis of HeLa cells labeling beta Actin with Mouse anti-beta Actin antibody (<a href="/products/EM21002" style="font-weight: bold;text-decoration: underline;">EM21002</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 Mouse anti-beta Actin antibody (<a href="/products/EM21002" style="font-weight: bold;text-decoration: underline;">EM21002</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 488, <a href="/products/HA1125" style="font-weight: bold;text-decoration: underline;">HA1125</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/ET1602-4" style="font-weight: bold;text-decoration: underline;">ET1602-4</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 594, <a href="/products/HA1122" style="font-weight: bold;text-decoration: underline;">HA1122</a>) was used as the secondary antibody at 1/1,000 dilution.

    Immunocytochemistry analysis of HeLa cells labeling beta Actin with Mouse anti-beta Actin antibody (EM21002) 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 Mouse anti-beta Actin antibody (EM21002) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) 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 (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) was used as the secondary antibody at 1/1,000 dilution.

  • Immunohistochemical analysis of paraffin-embedded mouse lung tissue with Mouse anti-beta Actin antibody (<a href="/products/EM21002" style="font-weight: bold;text-decoration: underline;">EM21002</a>) at 1/1,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/EM21002" style="font-weight: bold;text-decoration: underline;">EM21002</a>) at 1/1,000 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 lung tissue with Mouse anti-beta Actin antibody (EM21002) at 1/1,000 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 (EM21002) at 1/1,000 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 lung tissue with Mouse anti-beta Actin antibody (<a href="/products/EM21002" style="font-weight: bold;text-decoration: underline;">EM21002</a>) at 1/1,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/EM21002" style="font-weight: bold;text-decoration: underline;">EM21002</a>) at 1/1,000 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 lung tissue with Mouse anti-beta Actin antibody (EM21002) at 1/1,000 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 (EM21002) at 1/1,000 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.

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

Citation

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