beta Actin Mouse Monoclonal Antibody [A2-F6]

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Catalog# EM21002

beta Actin Mouse Monoclonal Antibody [A2-F6]

Application

WB
IF-Cell
IHC-P
FC

Reactivity

Human
Mouse
Rat

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

概述

产品名称

beta Actin Mouse Monoclonal Antibody [A2-F6]

抗体类型

Mouse Monoclonal Antibody

免疫原

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

种属反应性

Human, Mouse, Rat

验证应用

WB, IF-Cell, IHC-P, FC

分子量

Predicted band size: 42 kDa

阳性对照

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.

偶联

unconjugated

克隆号

A2-F6

RRID

AB_2819164

产品特性

形态

Liquid

浓度

2ug/ul

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

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

亚型

IgG1

纯化方式

Protein A affinity purified.

应用稀释度

WB
1:10,000-160,000

IF-Cell
1:100-1:200

IHC-P
1:1,000

FC
1:1,000

发表文章中的应用

WB	See 246 publications below
IHC	See 4 publications below
IF	See 3 publications below

发表文章中的种属

Mouse	See 69 publications below
Human	See 60 publications below
human	See 39 publications below

靶点

功能

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.

背景文献

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).

序列相似性

Belongs to the actin family.

翻译后修饰

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.

亚细胞定位

Cytoplasm.

UNIPROT #

SwissProt: P60709 Human

SwissProt: P60710 Mouse

SwissProt: P60711 Rat

别名

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

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图片

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 (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 (EM21002) at 1/20,000 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 1/20,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:150,000 dilution was used for 1 hour at room temperature.
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 (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 (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.