Anti-beta Actin antibody (ab8227)
Key features and details
- Rabbit polyclonal to beta Actin
- Suitable for: WB, IHC-P, ICC/IF
- Reacts with: Mouse, Rat, Rabbit, Chicken, Cow, Dog, Human, Xenopus laevis, Fish, Chinese hamster
- Isotype: IgG
Get better batch-to-batch reproducibility with a recombinant antibody
- Research with confidence – consistent and reproducible results with every batch
- Long-term and scalable supply – powered by recombinant technology for fast production
- Success from the first experiment – confirmed specificity through extensive validation
- Ethical standards compliant – production is animal-free
Overview
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Product name
Anti-beta Actin antibody
See all beta Actin primary antibodies -
Description
Rabbit polyclonal to beta Actin -
Host species
Rabbit -
Specificity
The immunogen used for this product shares 77% homology with Gamma actin/actin cytoplasmic 2. Cross-reactivity with this protein has not been confirmed experimentally. -
Tested applications
Suitable for: WB, IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Mouse, Rat, Rabbit, Chicken, Cow, Dog, Human, Xenopus laevis, Fish, Chinese hamster
Predicted to work with: Sheep, Guinea pig, Pig, Drosophila melanogaster, Monkey, Zebrafish, Rhesus monkey -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
(Peptide available asab13772) -
Positive control
- WB: A431, HeLa, Jurkat, HEK-293, NIH/3T3, MDCK, EBTr, SL-29, CHO and PC-12 whole cell lysate. Rat liver tissue lysate. HeLa nuclear lysate. Fish and rabbit liver. Xenopus laevis embryo. ICC: SV40LT-SMC and NIH/3T3 cells. IHC-P: Rat small intestine tissue. Human colon tissue.
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General notes
For western blot, milk blocking is suitable for use with fluorescent detection systems. For western blot using chemiluminescent (ECL) systems we recommend BSA blocking.
Abcam recommended secondaries - Goat Anti-Rabbit HRP (ab205718) and Goat Anti-Rabbit Alexa Fluor® 488 (ab150077).
See other anti-rabbit secondary antibodies that can be used with this antibody.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituents: PBS, 1% BSA
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help. -
Concentration information loading...
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Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Isotype control
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Recombinant Protein
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Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab8227 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB | (83) |
1/1000 - 1/5000.
We recommend Goat Anti-Rabbit IgG H&L (HRP) (ab6721) secondary antibody. A stronger signal is observed in chemiluminescent western blot when using 2% BSA as the blocking agent. Milk blocking is suitable for use with fluorescent detection systems. |
IHC-P | (2) |
Use a concentration of 0.2 - 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
We recommend Goat Anti-Rabbit IgG H&L (Biotin) (ab6720) secondary antibody or |
ICC/IF | (5) |
Use a concentration of 1 µg/ml.
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Notes |
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WB
1/1000 - 1/5000. We recommend Goat Anti-Rabbit IgG H&L (HRP) (ab6721) secondary antibody. A stronger signal is observed in chemiluminescent western blot when using 2% BSA as the blocking agent. Milk blocking is suitable for use with fluorescent detection systems. |
IHC-P
Use a concentration of 0.2 - 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. We recommend Goat Anti-Rabbit IgG H&L (Biotin) (ab6720) secondary antibody or |
ICC/IF
Use a concentration of 1 µg/ml. |
Target
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Function
Actins are highly conserved proteins that are involved in various types of cell motility and are ubiquitously expressed in all eukaryotic cells. -
Involvement in disease
Defects 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 similarities
Belongs to the actin family. -
Post-translational
modificationsISGylated. -
Cellular localization
Cytoplasm > cytoskeleton. Localized in cytoplasmic mRNP granules containing untranslated mRNAs. - Information by UniProt
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Database links
- Entrez Gene: 396526 Chicken
- Entrez Gene: 280979 Cow
- Entrez Gene: 403580 Dog
- Entrez Gene: 60 Human
- Entrez Gene: 11461 Mouse
- Entrez Gene: 414396 Pig
- Entrez Gene: 100009272 Rabbit
- Entrez Gene: 81822 Rat
see all -
Alternative names
- A26C1A antibody
- A26C1B antibody
- ACTB antibody
see all
Images
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All lanes : Anti-beta Actin antibody (ab8227) at 1/1000 dilution
Lane 1 : A431 (human epidermoid carcinoma cell line) Whole Cell Lysate
Lane 2 : HEK-293 (human epithelial cell line from embryonic kidney) Whole Cell Lysate
Lane 3 : NIH/3T3 (mouse embryo fibroblast cell line) Whole Cell Lysate
Lane 4 : PC-12 (rat adrenal gland pheochromocytoma cell line) Whole Cell Lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (Alexa Fluor® 790) (ab175781) at 1/10000 dilution
Observed band size: 42 kDa why is the actual band size different from the predicted?This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Milk before being incubated with ab8227 overnight at 4°C. Antibody binding was detected using Goat Anti-Rabbit IgG H&L (Alexa Fluor® 790) (ab175781) secondary antibody at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.
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ab8227 staining beta Actin in SV40LT-SMC (rat aortic smooth muscle cells transfected with SV40). The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab8227 at 1μg/ml (detected with ab150081, Alexa Fluor® 488 Goat anti-Rabbit, 1μg/ml, shown in green); and ab195889, Mouse monoclonal [DM1A] to alpha Tubulin - Microtubule Marker (Alexa Fluor® 594), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Actin antibody (ab8227)
IHC image of ab8227 staining beta Actin in rat small intestine formalin fixed paraffin embedded tissue sections, performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with EDTA (epitope retrieval solution 2) for 20 mins. The section was then incubated with ab8227, 0.2μg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. No primary antibody was used in the secondary only control (shown on the inset).
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times. -
ab8227 staining beta Actin in NIH/3T3 (mouse embryo fibroblast cell line) cells. The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab8227 at 1μg/ml (shown in green) and ab195889, Mouse monoclonal [DM1A] to alpha Tubulin - Microtubule Marker (Alexa Fluor® 594), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Actin antibody (ab8227)
IHC image of beta actin staining in a section of formalin-fixed paraffin-embedded normal human colon*. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 30mins. The section was then incubated with ab8227, 1/1000 dilution, for 15 mins at room temperature. A goat anti-rabbit biotinylated secondary antibody (ab6720, 1/1000 dilution) was used to detect the primary, and visualized using an HRP conjugated ABC system. Streptavidin HRP was used, ab7403 at a 1/10000 dilution. DAB was used as the chromogen (ab103723), diluted 1/100 and incubated for 10min at room temperature. The section was then counterstained with haematoxylin and mounted with DPX.
The inset negative control image is taken from an identical assay without primary antibody.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
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All lanes : Anti-beta Actin antibody (ab8227) at 1 µg/ml
Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate (blocked with 2% BSA)
Lane 2 : NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate (blocked with 2% BSA)
Lane 3 : Rat Liver tissue lysate (blocked with 2% BSA)
Lane 4 : HeLa whole cell lysate (blocked with 3% Milk)
Lane 5 : NIH/3T3 whole cell lysate (blocked with 3% Milk)
Lane 6 : Rat Liver tissue lysate (blocked with 3% Milk)
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) at 1/50000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Observed band size: 45 kDa why is the actual band size different from the predicted?
Exposure time: 10 secondsLanes 1-3: Blocked with 2% BSA
Lanes 4-6: Blocked with 3% Milk
This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin or 3% Milk before being incubated with ab8227 overnight at 4°C. Antibody binding was detected using an anti rabbit HRP secondary antibody, and visualised using ECL development solution ab133406
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All lanes : Anti-beta Actin antibody (ab8227) at 0.1 µg/ml
Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) Whole Cell Lysate
Lane 2 : NIH/3T3 (mouse embryonic fibroblast cell line) Whole Cell Lysate
Lane 3 : Liver (Rat) Tissue Lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Performed under reducing conditions.
Observed band size: 42 kDa why is the actual band size different from the predicted?
Exposure time: 1 minuteThis blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab8227 overnight at 4°C. Antibody binding was detected using an anti-rabbit HRP secondary antibody, and visualised using ECL development solution ab133406
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All lanes : Anti-beta Actin antibody (ab8227) at 1/1000 dilution
Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) nuclear lysate
Lane 2 : HeLa whole cell lysate
Lane 3 : A431 (human epidermoid carcinoma cell line) cell lysate
Lane 4 : Jurkat (human T cell leukemia cell line from peripheral blood) cell lysate
Lane 5 : HEK-293 (human epithelial cell line from embryonic kidney) cell lysate
Lane 6 : HeLa nuclear lysate withHuman beta Actin peptide (ab13772) at 1 µg/ml
Lane 7 : HeLa whole cell lysate withHuman beta Actin peptide (ab13772) at 1 µg/ml
Lane 8 : A431 cell lysate withHuman beta Actin peptide (ab13772) at 1 µg/ml
Lane 9 : Jurkat cell lysate withHuman beta Actin peptide (ab13772) at 1 µg/ml
Lane 10 : HEK-293 cell lysate withHuman beta Actin peptide (ab13772) at 1 µg/ml
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilution
Observed band size: 41.7 kDa why is the actual band size different from the predicted?
Exposure time: 5 seconds -
All lanes : Anti-beta Actin antibody (ab8227) at 1/1000 dilution
Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) cells
Lane 2 : NIH/3T3 (Mouse embryonic fibroblast cell line) cells
Lane 3 : Fish Liver
Lane 4 : Rabbit Liver
Lane 5 : MDCK (Canine kidney cell line) cells
Lane 6 : EBTr (cow trachea) cells
Lane 7 : SL-29 (chicken day 11 embryo) cells
Lane 8 : CHO (Chinese hamster ovary cell line) cells
Lane 9 : Xenopus laevis embryo
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab6721) at 1/5000 dilution
Observed band size: 41.7 kDa why is the actual band size different from the predicted?
Exposure time: 30 seconds -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Actin antibody (ab8227)
IHC image of beta Actin staining in normal human colon, formalin-fixed and paraffin-embedded tissue*. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 30mins. The section was incubated with ab8227, 3µg/ml overnight at +4°C. A anti-rabbit HRP secondary antibody (Ab97200, 1/200 dilution) was used for 1hr at room temperature. The section was counterstained with haematoxylin and mounted with DPX.
The inset negative control image is taken from an identical assay without primary antibody.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (3010)
ab8227 has been referenced in 3010 publications.
- Jiang R et al. METTL3 stabilizes HDAC5 mRNA in an m6A-dependent manner to facilitate malignant proliferation of osteosarcoma cells. Cell Death Discov 8:179 (2022). WB, IHC-P ; Human . PubMed: 35396379
- Chandler RJ et al. Promoterless, Nuclease-Free Genome Editing Confers a Growth Advantage for Corrected Hepatocytes in Mice With Methylmalonic Acidemia. Hepatology 73:2223-2237 (2021). PubMed: 32976669
- Goryszewska E et al. Pleiotropic role of prokineticin 1 in the porcine endometrium during pregnancy establishment and embryo implantation †. Biol Reprod 104:181-196 (2021). PubMed: 32997136
- Chen Z et al. Anti-inflammatory activity of Radix Angelicae biseratae in the treatment of osteoarthritis determined by systematic pharmacology and in vitro experiments. Exp Ther Med 21:5 (2021). PubMed: 33235614
- Liu Y et al. Inhibition of proliferation and migration and induction of apoptosis in glioma cells by silencing TLR4 expression levels via RNA interference. Oncol Lett 21:13 (2021). PubMed: 33240419