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Western blot - beta Actin antibody - Loading Control (ab8227)
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Immunocytochemistry/ Immunofluorescence - beta Actin antibody - Loading Control (ab8227)
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Western blot - beta Actin antibody - Loading Control (ab8227)
(enlarge)
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Product Name
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beta Actin antibody - 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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Rabbit polyclonal to beta Actin - Loading Control
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Immunogen
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Synthetic peptide 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, Chk, ChHm, Cow, Dog, Fsh, Mk, Pig, Rb, RMk, Shp, Xl
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Does not react with
(species key)
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Zfsh
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Tested applications
(see key)
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ELISA, ICC/IF, IHC-P, IP, WB
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Abreviews
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Application notes
(see key)
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Recommended dilutions
ELISA: 1:1000.
ICC/IF: Use at a concentration of 1 µg/ml.
IHC-P: Use at a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB: 1/1000 - 1/5000. Can be blocked with beta Actin peptide (ab13772).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
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Positive control
(see definition)
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Tested on HeLa whole cell lysate and mouse brain lysate
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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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Rabbit
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Clonality
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Polyclonal
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Isotype
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IgG
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Purity
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Immunogen affinity purified
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Storage buffer
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Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, 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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Batch dependent within range: 0.10 - 1.00 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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At Abcam, we have one centralized database to hold all of our product information, so that everything we know about this beta Actin antibody - 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 - Loading Control images, references, products related to ab8227 and other tools.
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beta Actin antibody - Loading Control images:
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 Western blot - beta Actin antibody - Loading Control (ab8227)
All lanes : beta Actin antibody - Loading Control (ab8227) at 1/5000 dilution
Lane 1 : HeLa whole cell extract
Lane 2 : Yeast cell extract
Lane 3 : Mouse brain tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
Goat polyclonal to Rabbit IgG - H&L (HRP) (ab6721) at 1/2000 dilution
Observed band size : 47 kDa (why is the actual band size different from the predicted?)
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Immunocytochemistry/ Immunofluorescence - beta Actin antibody - Loading Control (ab8227)
ICC/IF image of ab8227 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab8227, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - beta Actin antibody - Loading Control (ab8227)
IHC image of beta Actin staining in human liver FFPE section, performed on a BondTM system using the standard protocol F. 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 ab8227, 1µg/ml, for 8 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.
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Western blot - beta Actin antibody - Loading Control (ab8227)
All lanes : beta Actin antibody - Loading Control (ab8227) at 1/1000 dilution
Lane 1 : HeLa cells (Human) at 20 µg
Lane 2 : 3T3 cells (Mouse) at 20 µg
Lane 3 : Fish Liver at 20 µg
Lane 4 : Rabbit Liver at 20 µg
Lane 5 : MDCK cells (Dog) at 20 µg/ml
Lane 6 : EBTr cells (Cow) at 20 µg
Lane 7 : SL-29 cells (Chicken) at 20 µg/ml
Lane 8 : CHO cells (Chinese Hamster) at 20 µg
Lane 9 : Xenopus embryo at 20 µg
Secondary
Rabbit polyclonal to Goat IgG - H&L (HRP) (ab6741) at 1/5000 dilution
Observed band size : 41.7 kDa (why is the actual band size different from the predicted?)
Exposure time : 30 seconds
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References for beta Actin antibody - Loading Control (ab8227)
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This product has been used in: (two most recent references)
Mehedint MG et al. Choline deficiency alters global histone methylation and epigenetic marking at the Re1 site of the calbindin 1 gene. FASEB J 24:184-95 (2010). WB; Mouse. PubMed: 19752176
Kusinski LC et al. Isolation of plasma membrane vesicles from mouse placenta at term and measurement of system a and system beta amino acid transporter activity. Placenta 31:53-9 (2010). WB; Human, Mouse. PubMed: 19954844
See all 108 publication references for this product.
If you publish research using ab8227 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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