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Western blot - beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)
![Western blot - beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)](/ps/datasheet/images/8/ab8226/beta-Actin-Primary-antibodies-ab8226-27.jpg)
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Immunofluorescence - beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)
![Immunofluorescence - beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)](/ps/datasheet/images/ab8226_4.jpg)
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Western blot - beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)
![Western blot - beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)](/ps/datasheet/images/ab8226_3.jpg)
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Product Name
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beta Actin antibody [mAbcam 8226] - Loading Control
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See all beta Actin - Loading Control antibodies (8)...
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Product type
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Primary antibodies
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Description
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Mouse monoclonal [mAbcam 8226] to beta Actin - Loading Control
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Immunogen
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Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human beta Actin.
(Peptide available as ab13772.)
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Reacts with
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Hu, Ms, Rat, AGMk, ArHm, Chk, ChHm, Cow, Dog, Pig, Rb
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Predicted to react
(species key)
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Gpig, Hrs, Shp (Due to sequence homology)
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Specificity
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Does not cross-react with adult cardiac, smooth, or skeletal muscle actin.
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Tested applications
(see key)
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Flow Cyt, ICC/IF, IP, WB
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Abreviews
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Application notes
(see key)
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Recommended dilutions Flow Cyt: Use at an assay dependent dilution. ICC/IF: Use at a concentration of 5 µg/ml. IP: 1:100 WB: 1/500 - 1/10000. Predicted molecular weight: 42 kDa. Can be blocked with beta Actin peptide (ab13772).
Abcam recommends blocking and incubation using 5% BSA as we have found that use of 5% milk significantly reduces the intensity of the signal obtained. Please refer to WB image for further details.
Not yet tested in other applications. Optimal dilutions/concentrations should be determined by the end user.
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Cellular localization
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Cytoplasmic
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Research areas
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Signal Transduction >> Cytoskeleton / ECM >> Cytoskeleton >> Microfilaments >> Actin etc >> Actin Tags & Cell Markers >> Subcellular Markers >> Cytoskeleton >> Actin Isotype/Loading Controls >> Loading Controls >> Beta Actin
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Relevance
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Beta actin is one of six different actin isoforms which have been identified. Actins are highly conserved proteins that are involved in cell motility, structure and integrity. Be aware when using beta actin antibodies as loading controls that although beta actin is normally the best protein to use as a loading control, under some conditions its amounts can vary between samples. In particular, expression of actin in adipose tissue is very low (Spiegelman and Farmer, Cell, 1982, 29(1):53-60) "in cells undergoing adipose differentiation actin synthesis decreases by 90%".
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Database links
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The links below go to external sites and will open in a new browser window
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Raised in
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Mouse
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Clonality
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Monoclonal
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Clone number
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mAbcam 8226
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Myeloma
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Sp2/0-Ag14
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Isotype
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IgG1
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Light chain type
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kappa
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Purity
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Protein G purified
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Storage buffer
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Preservative: 0.02% Sodium Azide Constituents: PBS, pH 7.4 Material safety datasheet (MSDS) for this product: Sodium Azide MSDS
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Form
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Liquid
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Concentration
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1.000 mg/ml
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Storage instructions
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Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
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Notes
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The concentration of this antibody is batch dependent.
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At Abcam, we have one centralized database to hold all of our product information, so that everything we know about this beta Actin antibody [mAbcam 8226] - Loading Control is on this datasheet. But please do contact us if you would like any reassurance! |
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See below for beta Actin antibody [mAbcam 8226] - Loading Control images, references, products related to ab8226 and other tools.
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beta Actin antibody [mAbcam 8226] - Loading Control images:
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Western blot - beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)
Lanes 1 - 3 : beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1 µg/ml (5% BSA BLOCK) Lanes 4 - 6 : beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1 µg/ml (5% MILK BLOCK)
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate Lane 3 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate Lane 4 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate Lane 5 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate Lane 6 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
Lysates/proteins at 10 µg per lane.
Secondary 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
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Immunofluorescence - beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)
Immunofluorescence using ab8226 at 5ug/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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Western blot - beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)
beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 0.5 µg/ml + HeLa cell lysate
Secondary Goat polyclonal to mouse IgG H&L (HRP) at 1/5000 dilution
Performed under non-reducing conditions.
Predicted band size : 42 kDa Observed band size : 42 kDa
Exposure time : 30 seconds
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Immunocytochemistry - beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)
ab8226 staining beta Actin in human AGS cells by Immunocytochemistry. Cells were fixed with formaldehyde, permeabilized with 0.025% Triton-X in TBS and blocking with 5% serum was performed for 1 hour at 230C. Samples were incubated with primary antibody (1/500) for 1 hour at 23°C. An HRP®-conjugated goat polyclonal to mouse IgG was used as secondary antibody. This image is a courtesy of Anonimous Abreview
See Abreview
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Western blot - beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)
Lane 1 : beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1/1000 dilution Lane 2 : beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1/10000 dilution Lanes 3 - 4 : beta Actin antibody [mAbcam 8226] - Loading Control (ab8226) at 1/500 dilution
Lane 1 : HeLa Cell lysate Lane 2 : HeLa Cell lysate Lane 3 : 293 Cell lysate Lane 4 : 3T3 Mouse Cell lysate
Lysates/proteins at 20 µg per lane.
Secondary Rabbit polyclonal to Mouse IgG - H&L (HRP) (ab6728) at 1/5000 dilution
Performed under reducing conditions.
Predicted band size : 42 kDa
Exposure time : 10 seconds
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References for beta Actin antibody [mAbcam 8226] - Loading Control (ab8226)
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This product has been used in: (two most recent references)
Xu K et al. Lunatic Fringe-mediated Notch signaling is required for lung alveogenesis. Am J Physiol Lung Cell Mol Physiol 298:L45-56 (2010). WB; Mouse. PubMed: 19897741
Vreede FT et al. Mechanisms and functional implications of the degradation of host RNA polymerase II in influenza virus infected cells. Virology 396:125-34 (2010). WB; Human. PubMed: 19875144
See all 78 publication references for this product.
If you publish research using ab8226 please let us know so that we can cite the reference on this datasheet.
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Search PubMed (MEDLINE) for references to beta Actin
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beta Actin antibody [mAbcam 8226] - Loading Control - more information
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