Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)
Key features and details
- Mouse monoclonal [mAbcam 8226] to beta Actin - Loading Control
- Suitable for: ICC/IF, IHC-P, WB
- Reacts with: Mouse, Rat, Human
- Isotype: IgG1
Related conjugates and formulations
Overview
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Product name
Anti-beta Actin antibody [mAbcam 8226] - Loading Control
See all beta Actin primary antibodies -
Description
Mouse monoclonal [mAbcam 8226] to beta Actin - Loading Control -
Host species
Mouse -
Specificity
Does not cross-react with adult cardiac, smooth, or skeletal muscle actin. 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: ICC/IF, IHC-P, WBmore details -
Species reactivity
Reacts with: Mouse, Rat, Human
Predicted to work with: Sheep, Rabbit, Horse, Chicken, Guinea pig, Cow, Dog, Pig, Monkey, Zebrafish, African green monkey, Chinese hamster, Armenian hamster -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
(Peptide available asab13772) -
Positive control
- WB, ICC/IF: HeLa, Jurkat, A431, HEK293, NIH 3T3, PC12 cells. IHC-P: Human colon (FFPE), Rat colon (FFPE) tissue. Hela cells
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General notes
Western blot protocol advice:
We recommend blocking with 2-5% BSA as we have found that use of 5% milk significantly reduces the band intensity for beta actin. Please see the comparison data in the images section. If milk block is required, we recommend using ab8224 mouse monoclonal [mAbcam 8224] to beta actin. Contact our Scientific Support team for more information or advice.
This antibody clone [mAbcam 8226] is manufactured by Abcam.
If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.
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, 6.97% L-Arginine -
Concentration information loading...
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Purity
Protein G purified -
Clonality
Monoclonal -
Clone number
mAbcam 8226 -
Myeloma
Sp2/0-Ag14 -
Isotype
IgG1 -
Light chain type
kappa -
Research areas
Associated products
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Alternative Versions
- Alexa Fluor® 680 Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab184092)
- Alexa Fluor® 790 Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab184576)
- HRP Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab20272)
- Anti-Actin antibody [mAbGEa] (ab230169)
- Anti-beta Actin antibody [mAbcam 8226] - BSA and Azide free (ab264083)
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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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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab8226 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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ICC/IF | (17) |
Use a concentration of 5 µg/ml.
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IHC-P | (1) |
Use a concentration of 0.1 - 0.5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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WB | (100) |
Use a concentration of 1 µg/ml. Predicted molecular weight: 42 kDa.Can be blocked with Human beta Actin peptide (ab13772).
We recommend blocking using 2-5% BSA as we have found that use of 5% milk significantly reduces the band intensity for beta actin. Please refer to the images section for the blocking comparison data. |
Notes |
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ICC/IF
Use a concentration of 5 µg/ml. |
IHC-P
Use a concentration of 0.1 - 0.5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
WB
Use a concentration of 1 µg/ml. Predicted molecular weight: 42 kDa.Can be blocked with Human beta Actin peptide (ab13772). We recommend blocking using 2-5% BSA as we have found that use of 5% milk significantly reduces the band intensity for beta actin. Please refer to the images section for the blocking comparison data. |
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: 100033878 Horse
- Entrez Gene: 60 Human
- Entrez Gene: 11461 Mouse
- Entrez Gene: 414396 Pig
- Entrez Gene: 100009272 Rabbit
see all -
Alternative names
- A26C1A antibody
- A26C1B antibody
- ACTB antibody
see all
Images
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Immunocytochemistry/ Immunofluorescence - Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)
ab8226 staining Beta Actin in HeLa cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-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 ab8226 at 5µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150117, Goat polyclonal Secondary Antibody to Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).
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All lanes : Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1/1000 dilution
Lane 1 : A431 (Human epidermoid carcinoma cell line) Whole Cell Lysate
Lane 2 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lane 3 : NIH 3T3 (Mouse embryo fibroblast cell line) Whole Cell Lysate
Lane 4 : PC12 (Rat adrenal gland pheochromocytoma cell line) Whole Cell Lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Mouse IgG H&L (Alexa Fluor® 790) (ab175783) at 1/10000 dilution
Predicted band size: 42 kDa
Observed band size: 42 kDaThis 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 ab8226 overnight at 4°C. Antibody binding was detected using Goat Anti-Mouse IgG H&L (Alexa Fluor® 790) (ab175783) at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.
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Immunocytochemistry/ Immunofluorescence - Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)This image is a courtesy of Anonymous Abreview
ab8226 staining beta Actin in human gastric epithelial AGS cells by Immunocytochemistry/ Immunofluorescence. Cells were fixed in formaldehyde. Permeabilization and blocking was carried out using 5% BSA containing 0.025% Triton X in TBS for 1 hour at 23°C. Primary antibody was used at 5µg/ml for 1 hour at 23°C. An Alexa Fluor 488® conjugated goat polyclonal to mouse IgG was used as the secondary antibody at a 1/1000 dilution.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)
IHC image of ab8226 staining beta Actin in human colon formalin fixed paraffin embedded tissue sections*, performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab8226, 0.1μg/ml working concentration, 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.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre -
Immunocytochemistry/ Immunofluorescence - Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)
Immunofluorescence using ab8226 at 5µg/ml incubated for 1 hour on Rat Colon Cancer cells.
Cells were fixed with ice-cold methanol for 5 mins, then for all following steps, permeabilised in TBS-T for 30 mins, blocked with 5% BSA for 30 mins and then washed in TBS-T. Secondary antibody was Alexa Fluor 488 goat anti-mouse IgG at 1/1000 incubated for 1 hour. Cells were counterstained with DAPI. Image at 400X magnification. All incubations were at room temperature.
The beta actin fibres can be seen arrayed around the edge of the cells.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)
IHC image of ab8226 staining beta Actin in rat colon formalin fixed paraffin embedded tissue sections, performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab8226, 0.5μg/ml working concentration, 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. -
Lane 1 : Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1/1000 dilution
Lane 2 : Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1/10000 dilution
Lanes 3-4 : Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1/500 dilution
Lanes 1-2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate
Lane 3 : HEK293 (Human epithelial cell line from embryonic kidney) cell lysate
Lane 4 : NIH/3T3 (Mouse embryo fibroblast cell line) cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Rabbit Anti-Mouse IgG H&L (HRP) (ab6728) at 1/5000 dilution
Performed under reducing conditions.
Predicted band size: 42 kDa
Exposure time: 10 seconds -
All lanes : Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1 µg/ml
Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Lane 3 : A431 (Human epidermoid carcinoma cell line) whole cell lysate
Lane 4 : HEK293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 5 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : HRP-conjugated goat anti-mouse IgG at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 42 kDa
Observed band size: 42 kDa
Exposure time: 10 secondsWestern blot image using the Optiblot Reducing Electrophoresis Kit - 10 x 10 cm (4-20%) with the Prism Ultra Protein Ladder (ab116028) 5µl used. We recommend using our ECL substrate kit (ab65623).
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Lanes 1-3 : Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1 µg/ml (5% BSA BLOCK)
Lanes 4-6 : Anti-beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1 µg/ml (5% MILK BLOCK)
Lanes 1 & 4 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lanes 2 & 5 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Lanes 3 & 6 : NIH/3T3 (Mouse embryo fibroblast cell line) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
Performed under reducing conditions.
Predicted band size: 42 kDa
Observed band size: 42 kDa
Exposure time: 8 minutes
Datasheets and documents
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SDS download
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Datasheet download
References (2899)
ab8226 has been referenced in 2899 publications.
- Wang S et al. Knockdown of circ_0004585 enhances the chemosensitivity of colorectal cancer cells to 5-fluorouracil via the miR-874-3p/CCND1 axis. Histol Histopathol 38:99-112 (2023). PubMed: 35900059
- Liu Q et al. PUM2 aggravates the neuroinflammation and brain damage induced by ischemia-reperfusion through the SLC7A11-dependent inhibition of ferroptosis via suppressing the SIRT1. Mol Cell Biochem 478:609-620 (2023). PubMed: 35997855
- Fang M et al. MicroRNA-29b regulates pyroptosis involving calcific aortic valve disease through the STAT3/SOCS1 pathway. Int J Cardiol 371:319-328 (2023). PubMed: 36064035
- Li Y et al. Enhancing cartilage repair with optimized supramolecular hydrogel-based scaffold and pulsed electromagnetic field. Bioact Mater 22:312-324 (2023). PubMed: 36263100
- Zheng Y et al. LncNAP1L6 activates MMP pathway by stabilizing the m6A-modified NAP1L2 to promote malignant progression in prostate cancer. Cancer Gene Ther 30:209-218 (2023). PubMed: 36195720