Invitrogen

beta Actin Monoclonal Antibody (15G5A11/E2)

Advanced Verification

This Antibody was verified by Cell treatment to ensure that the antibody binds to the antigen stated. View Details

3 Published Figures

7 References

View all (48) beta Actin antibodies

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Datasheet

Questions & Answers

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beta Actin Antibody in Western Blot (WB) — Western blot analysis of beta-actin was performed by loading 25 µg of various cell lysates and 10 µL of PageRuler Prestained Protein Ladder (Product # 26616) per well onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked with StartingBlock T20 (TBS) Blocking Buffer (Product # 37543). The membrane was probed with a beta-actin monoclonal antibody (Product # MA1-140) at a dilution of 1:16,000 overnight at 4°C on a rocking platform, washed in TBS-0.1%Tween-20, and probed with a goat anti-mouse IgG-HRP secondary antibody (Product # 31430) at a dilution of 1:40,000 for at least 1 hour. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34080). NOTE: Similar results are obtained when blocking with 5% BSA in TBST.

beta Actin Antibody in Immunofluorescence (IF) — Immunofluorescent analysis of Beta- Actin (green) in U2OS cells untreated or treated with 1uM Actinomycin D for 20 hours. The cells were fixed with 4% Paraformaldehyde for 15 minutes, permeabilized with 0.1% Triton X-100 for 15 minutes, and blocked with 3% BSA for 30 minutes at room temperature. Cells were stained with a Beta-Actin mouse monoclonal antibody (Product # MA1-140) at a dilution of 1:2000 in blocking buffer for 1 hour at room temperature, and then incubated with a Goat anti-Mouse IgG (H+L) Secondary Antibody, Alexa Fluor Plus 488 conjugate (Product # A32723) at a dilution of 1:500 for at least 30 minutes at a room temperature in the dark (green). Nuclei (blue) were stained with Hoechst 33342 (Product # 62249). Images were taken on a Thermo Scientific ToxInsight Instrument at 20X magnification.

beta Actin Antibody in Immunoprecipitation (IP) — Immunoprecipitation of beta-actin was performed using NIH/3T3 cells. Antigen-antibody complexes were formed by incubating 500 µg of NIH/3T3 whole cell lysates with 2 µg of a beta-actin monoclonal antibody (Product # MA1-140) overnight on a rocking platform at 4°C. The immune complexes were captured on 100 µL Protein A/G Agarose (Product # 20421), washed extensively, and eluted with 5X Lane Marker Reducing Sample Buffer (Product # 39000). The IP eluate (right lane) and 15 µg of NIH/3T3 lysate (left lane) were resolved on a 4-20% Tris-HCl polyacrylamide gel, transferred to a PVDF membrane, and blocked with 5% BSA/TBS-0.1%Tween for at least 1 hour. The membrane was probed with a beta-actin monoclonal antibody (Product # MA1-140) at a dilution of 1:8000 overnight rotating at 4°C, washed in TBST, and probed with n HRP-conjugated anti-mouse light chain secondary antibody a dilution of 1:40,000 for at least 1 hour. Chemiluminescent detection was performed using SuperSignal West Pico (Product # 34080).

beta Actin Antibody in Cell treatment — The specificity of anti-Beta-Actin mouse monoclonal antibody (Product # MA1-140) was demonstrated by the immunofluorescence detection of the disruption of actin filaments in U-2OS cells upon treatment with Actinomycin D (upper panel) compared to untreated cells (bottom panel). {TM}

beta Actin Antibody — Figure 2. Generation of the CRISPR/Cas9-based lentivirus and HIF-1 alpha knockout in SMMC-7721 cells and in the SMMC-7721-induced tumor tissues of mice. (A) Immunohistochemical analysis for the detection of HIF-1alpha expression in hepatocellular carcinoma tissues following infection with LV-Ctrl or LV-H721 in the subcutaneous animal model (bars, 100 um). (B) Diagram illustration of the lentiviral vector ( Lenti-CAS9-sgRNA-egfp ). Three sgRNAs (sgRNA719, sgRNA720 and sgRNA721) targeting HIF-1alpha were designed, and Lenti-CAS9-sgRNA719 / 720 /721 plasmids were constructed. (C) Gel electrophoresis and DNA analysis of the HIF-1alpha genomic frame shift mutation conducted after the T7E1 endonuclease assay in SMMC-7721 cells infected with LV-Ctrl or LV-H719/720/721. (D) Western blot analysis of HIF-1alpha expression in the different lentivirus-infected SMMC-7721 cells with CoCl 2 -simulated hypoxia. (E) GFP-positive cells analyzed by flow cytometry following infection with LV-Ctrl and LV-H721. (F) Relative mRNA and (G) protein expression levels of VEGF and MDR1 in different cell groups were examined by reverse transcription-quantitative polymerase chain reaction and western blot analysis. beta-actin served as an internal control. Data are a representation of four repeated experiments, and histograms represent the mean +- standard deviation. ***P<0.001. sgRNA, small guide RNA; Ctrl, control; LV, lentivirus; HIF-1alpha, hypoxia inducible factor-1alpha; VEGF, vascular endotheli

Product Details

Applications

Tested Dilution

Publications

Western Blot (WB)

1:5000-1:20,000

View 7 publications 7 publications

Immunocytochemistry (ICC)

1:1000-1:4000

Immunofluorescence (IF)

1:1000-1:4000

Immunoprecipitation (IP)

2 µg

Product Specifications

Species Reactivity

Dog, Human, Mouse, Non-human primate, Rat

Published species

Human, Mouse, Rat

Host/Isotype

Mouse / IgG1

Class

Monoclonal

Type

Antibody

Clone

15G5A11/E2

Immunogen

Purified full-length recombinant human beta-actin expressed in E. coli

Conjugate

Unconjugated

Form

Liquid

Concentration

1 mg/mL

Purification

Protein A

Storage buffer

PBS with 1mg/mL BSA, 30% glycerol

Contains

0.05% sodium azide

Storage conditions

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

RRID

AB_2536844

Product Specific Information

By Western blot, MA1-140 detects a prominent ~42kD protein in human, mouse, non-human primate, and rat whole cell lysates.

For visualization of actin filaments by immunofluorescence, fixation and permeabilization with methanol is required.

Target Information

Beta actin is one of six actin isoforms which have been identified. Beta actin is a non-muscle cytoskeletal protein in all human cell types and is involved in cell motility, structure, and integrity. There are six different actin isoforms in human. The beta isoform of actin, along with gamma actin, coexist in most cell types as components of the cyto-skeleton. Beta actins are cytoplasmic proteins ubuquitously expressed in all eukaryotic cells. Polymerization of globular actin (G-actin) leads to a structural filament (F-actin) in the form of two stranded helix. Actins are highly conserved proteins that are involved in cell motility, structure and integrity. Because beta actin is ubiquitously expressed in all eukaryotic cells, it is frequently used as a loading control for assays involving protein detection, such as Western blotting. Antibodies to beta-actin provide a specific and useful tool in studying the intracellular distribution of beta-actin and the static and dynamic aspects of the cytoskeleton.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Protein Aliases: actin, beta; Actin, cytoplasmic 1; Actin, cytoplasmic 1, N-terminally processed; Atcb; beta cytoskeletal actin; Beta-actin; Cap Z; capZ beta; cytoplasmic beta-actin; cytoskeleton actin; gamma actin; PS1TP5-binding protein 1

Gene Aliases: ACTB; Actx; beta-actin; BRWS1; E430023M04Rik; PS1TP5BP1

UniProt ID: (Human) P60709, (Rat) P60711, (Mouse) P60710

Entrez Gene ID: (Human) 60, (Dog) 403580, (Rat) 81822, (Mouse) 11461

Molecular Function: actin and actin related protein actin family cytoskeletal protein cytoskeletal protein