PRODUCT CODE: EM21002

Beta Actin Mouse Monoclonal Antibody [A2-F6] (EM21002)

Applications

  • WB

  • ICC

  • IHC-P

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

Western blot analysis of Beta Actin on Hela cell lysates with Mouse anti-Beta Actin antibody (EM21002).<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 (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 Hela cell lysates with Mouse anti-Beta Actin antibody (EM21002).<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 (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.<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 (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. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (EM21002, 1/10,000) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:20,000 dilution was used for 1 hour at room temperature.<br />
<br />
Positive control:<br />
Lane 1: NIH/3T3 cell lysate, 10 µg/Lane<br />
Lane 2: PC-12 cell lysate, 10 µg/Lane<br />
Lane 3: MCF-7 cell lysate, 10 µg/Lane<br />
Lane 4: HepG2 cell lysate, 10 µg/Lane<br />
Lane 5: Hela cell lysate, 10 µg/Lane<br />
Lane 6: Mouse lung tissue lysate, 20 µg/Lane
  • ICC staining of Beta Actin in Hela cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (EM21002, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 conjugate-Goat anti-Mouse IgG was used as the secondary antibody at 1/1,000 dilution.
  • ICC staining of Beta Actin in A549 cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (EM21002, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 conjugate-Goat anti-Mouse IgG was used as the secondary antibody at 1/1,000 dilution.
  • ICC staining of Beta Actin in NIH/3T3 cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (EM21002, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®594 conjugate-Goat anti-Mouse IgG was used as the secondary antibody at 1/1,000 dilution.
  • Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue using anti-Beta Actin antibody. 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 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM21002, 1/50) for 30 minutes 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 prostate tissue using anti-Beta Actin antibody. 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 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM21002, 1/50) for 30 minutes 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 kidney tissue using anti-Beta Actin antibody. 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 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (EM21002, 1/50) for 30 minutes 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.
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.

Applications

  • WB

  • ICC

  • IHC-P

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

SPECIFICATIONS

Product Type

Mouse monoclonal primary

Product Name

Beta Actin Mouse Monoclonal Antibody [A2-F6] (EM21002)

Immunogen

Synthetic peptide (klh-coupled) within human beta-actin n terminal.

Host

Mouse

Positive Control

NIH/3T3 cell lysate, PC-12 cell lysate, MCF-7 cell lysate, HepG2 cell lysate, Hela cell lysate, mouse lung tissue lysate, Hela, A549, NIH/3T3, human colon carcinoma tissue, mouse prostate tissue, mouse kidney tissue.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

A2-F6

PROPERTIES

Form

Liquid

Storage Condition

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

Storage Buffer

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

Concentration

2ug/ul

PURIFICATION

Protein A affinity purified.

MOLECULAR WEIGHT

42 kDa

Isotype

IgG1

APPLICATION DILUTION

  • WB

  • 1:10,000-160,000

  • ICC

  • 1:100-1:200

  • IHC-P

  • 1:100-1:200

  • FC

  • 1:100-1:200

TARGET

UNIPROT #

PROTEIN NAME

Beta Actin

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;Beta-actin antibody;BRWS1 antibody;E430023M04Rik antibody;MGC128179 antibody;PS1TP5 binding protein 1 antibody;PS1TP5BP1 antibody

SEQUENCE SIMILARITIES

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.

FUNCTION

Beta-actin 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 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.

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