Key features and details
- Mouse monoclonal [AC-15] to beta Actin
- Suitable for: ICC/IF, WB
- Knockout validated
- Reacts with: Mouse, Rat, Sheep, Rabbit, Chicken, Guinea pig, Hamster, Cow, Cat, Dog, Carp, African green monkey, Opossum, Chinese hamster, Common marmoset, Meriones unguiculatus
- Isotype: IgG1
Product nameAnti-beta Actin antibody [AC-15]
See all beta Actin primary antibodies
DescriptionMouse monoclonal [AC-15] to beta Actin
Tested applicationsSuitable for: ICC/IF, WBmore details
Species reactivityReacts with: Mouse, Rat, Sheep, Rabbit, Chicken, Guinea pig, Hamster, Cow, Cat, Dog, Carp, African green monkey, Opossum, Chinese hamster, Common marmoset, Meriones unguiculatus
Does not react with: Drosophila melanogaster, Dictyostelium discoideum
Synthetic peptide corresponding to beta Actin aa 1-14 (N terminal) conjugated to Keyhole Limpet Haemocyanin (KLH). Slightly modified ?-cytoplasmic actin N-terminal peptide, Ac-Asp-Asp-Asp-Ile-Ala-Al?a-Leu-Val-Ile-Asp-Asn-Gl y?-Ser-Gly-Lys, conjugated to KLH.
EpitopeN-terminal of the beta isoform of actin.
- ICC/IF: SV40LT-SMC cells WB: HAP1, HeLa, Jurkat, A431, HEK-293, COS-7, NIH/3T3, PC-12, Rat2, CHO, MDBK and MDCK cell lysates. IHC-Fr: Human stomach tissue. ICC/IF: SV40LT-SMC cells.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage bufferpH: 7.40
Preservative: 0.097% Sodium azide
Concentration information loading...
Purification notesPurified from hybridoma cell culture.
- Prestained Protein Ladder - Broad molecular weight (10-245 kDa) (ab116028)
- Human beta Actin peptide (ab13772)
- Phalloidin-iFluor 488 Reagent (ab176753)
- Phalloidin-iFluor 647 Reagent (ab176759)
- Anti-beta Actin antibody [mAbcam 8224] - Loading Control (HRP) (ab197277)
- Anti-beta Actin antibody [mAbcam 8226] - Loading Control (HRP) (ab20272)
- Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291)
- Anti-beta Actin antibody [mAbcam 8224] - Loading Control (ab8224)
- Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)
Our Abpromise guarantee covers the use of ab6276 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use a concentration of 5 μg/ml.|
|WB||1/5000 - 1/16000. Predicted molecular weight: 42 kDa.|
FunctionActins are highly conserved proteins that are involved in various types of cell motility and are ubiquitously expressed in all eukaryotic cells.
Involvement in diseaseDefects in ACTB are a cause of dystonia juvenile-onset (DYTJ) [MIM:607371]. DYTJ is a form of dystonia with juvenile onset. Dystonia is defined by the presence of sustained involuntary muscle contraction, often leading to abnormal postures. DYTJ patients manifest progressive, generalized, dopa-unresponsive dystonia, developmental malformations and sensory hearing loss.
Sequence similaritiesBelongs to the actin family.
Cellular localizationCytoplasm > cytoskeleton. Localized in cytoplasmic mRNP granules containing untranslated mRNAs.
- Information by UniProt
- A26C1A antibody
- A26C1B antibody
- ACTB antibody
Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: Beta actin knockout HAP1 cell lysate (20 µg)
Lanes 1 and 2: Merged signal (red and green). Green - beta actin, ab6276 observed at 42 kDa. Red - loading control, ab181602 observed at 37 kDa.
Ab6276 was shown to specifically react with beta actin in wild-type HAP1 cells. No band was observed when beta actin knockout samples were used. Wild-type and beta actin knockout samples were subjected to SDS-PAGE. ab6276 (beta actin) and ab181602 (loading control to GAPDH) were diluted 1/5000 and 1/10 000 and incubated overnight at 4°C. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.
ab6276 staining beta Actin in SV40LT-SMC cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized in 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab6276 at a working concentration of 5μg/ml and ab190573, Rabbit monoclonal [EP1332Y] to alpha Tubulin (Alexa Fluor® 647, shown in red) at 1/250 overnight at +4°C, followed by a further incubation at room temperature for 1h with an anti-mouse AlexaFluor® 488 (ab150117) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Western Blot of ab6276 (used as loading control) with whole tissue lysate of grey matter from BA20 (temporal cortex). Ab6276 was diluted 1/50000 and incubated with the sample for 16 hours at 4°C. 5 µg of lysate was loaded onto the gel, which was blocked with 5% milk for 1 hour at 20°C. An Alexa Fluor® 680 conjugated goat anti-mouse antibody, diluted 1/5000, was used as the secondary.
Bands below actin in image are synaptophysin (SYN).
All lanes : Anti-beta Actin antibody [AC-15] (ab6276) at 1 μg/ml
Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) cell lysate
Lane 2 : Jurkat (human T cell leukemia cell line from peripheral blood) cell lysate
Lane 3 : COS-7 (african green monkey kidney fibroblast-like cell line) cell lysate
Lane 4 : NIH/3T3 (mouse embryonic fibroblast cell line) cell lysate
Lane 5 : PC-12 (rat adrenal gland pheochromocytoma cell line) cell lysate
Lane 6 : Rat2 (rat fibroblast cell line) cell lysate
Lane 7 : CHO (chinese hamster ovary cell line) cell lysate
Lane 8 : MDBK (bovine kidney cell line) cell lysate
Lane 9 : MDCK (canine kidney cell line) cell lysate
All lanes : Goat Anti-Mouse IgG-Peroxidase
Developed using the ECL technique.
Predicted band size: 42 kDa
All lanes : Anti-beta Actin antibody [AC-15] (ab6276) at 1/5000 dilution
Lane 1 : HeLa nuclear
Lane 2 : HeLa whole cell lysate
Lane 3 : A431 cell lysate
Lane 4 : Jurkat cell lysate
Lane 5 : HEK293 cell lysate
Lysates/proteins at 20 μg per lane.
All lanes : Alexa Fluor anti mouse at 1/5000 dilution
Performed under reducing conditions.
Predicted band size: 42 kDa
Observed band size: 42 kDa
MDCK cells induced with increasing amounts of doxycycline to control expression of the gene of interest. All cells were normalized for loading with an albumin protein standard assay. Anti-beta actin (ab6276) was used at a concentration of 1:5000 in a milk blocking solution. B-actin blotting confirms the albumin assay in showing that an equal amount of lysate was loaded in each lane.
MDCK cells induced with increasing amounts of doxycycline to control expression of the gene of interest. All cells were normalized for loading with an albumin protein standard asssay. Anti-beta actin (ab6276) was used at a concentration of 1:5000 in a milk blocking solution. B-actin blotting confirms the albumin assay in showing that an equal amount of lysate was loaded in each lane.
ab6276 has been referenced in 1600 publications.
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- Dong Y et al. Schizandrin A Protects Human Retinal Pigment Epithelial Cell Line ARPE-19 against HG-Induced Cell Injury by Regulation of miR-145. Mol Ther Nucleic Acids 19:42-49 (2020). PubMed: 31794890
- Pungsrinont T et al. Senolytic compounds control a distinct fate of androgen receptor agonist- and antagonist-induced cellular senescent LNCaP prostate cancer cells. Cell Biosci 10:59 (2020). PubMed: 32351687
- Han X et al. Advanced glycation end products enhance macrophage polarization to the M1 phenotype via the HIF-1a/PDK4 pathway. Mol Cell Endocrinol 514:110878 (2020). PubMed: 32464167
- Akdag M et al. Proximal Biotinylation-Based Combinatory Approach for Isolating Integral Plasma Membrane Proteins. J Proteome Res N/A:N/A (2020). PubMed: 32500712