Anti-beta Actin antibody [mAbcam 8224] - Loading Control (ab8224)

Overview

• Product nameAnti-beta Actin antibody [mAbcam 8224] - Loading ControlSee all beta Actin primary antibodies ...
• Description
  Mouse monoclonal [mAbcam 8224] to beta Actin - Loading Control
• SpecificityRecognises a single band at 42kD representing beta Actin.
• Tested applicationsFlow Cyt, IHC-P, WB, ICC/IF more details
• Species reactivity
  Reacts with: Mouse, Rat, Rabbit, Chicken, Cow, Cat, Dog, Human, Pig, Saccharomyces cerevisiae, Xenopus laevis, Fruit fly (Drosophila melanogaster), Schizosaccharomyces pombe, Chinese Hamster, Other
  
• Immunogen

  Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human beta Actin.

  Read Abcam's proprietary immunogen policy

  (Peptide available as ab13772.)

• Positive controlIn Flow Cytometry, this antibody gave a positive signal in methanol fixed/Tween permeabilised HeLa cells.
• General notes

  
  

  This clone works well as a loading control for Xenopus, Drosophila, S. cerevisiae and S.pombe. We recommend using ab8224 instead of ab8226 for these species.

Properties

• FormLiquid
• Storage instructionsStore at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
• Storage bufferPreservative: 0.02% Sodium Azide
  Constituents: Tris HCl, 500mM Sodium chloride, 200mM Sodium citrate. pH 7.5
• Concentration information loading...
• PurityIgG fraction
• Primary antibody notes This clone works well as a loading control for Xenopus, Drosophila, S. cerevisiae and S.pombe. We recommend using ab8224 instead of ab8226 for these species.
• Clonality Monoclonal
• Clone numbermAbcam 8224
• MyelomaSp2/0-Ag14
• IsotypeIgG3
• Light chain typekappa
• Research Areas
  • Signal Transduction
  • Cytoskeleton / ECM
  • Cytoskeleton
  • Intermediate Filaments
  • Class I
  • Cytokeratins

  • Signal Transduction
  • Cytoskeleton / ECM
  • Cytoskeleton
  • Microfilaments
  • Actin etc
  • Actin

  • Isotype/Loading Controls
  • Loading Controls
  • Beta Actin

  • Tags & Cell Markers
  • Subcellular Markers
  • Cytoskeleton
  • Actin

Applications

Target

• 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.
• Post-translational
  modificationsISGylated.
• Cellular localizationCytoplasm > cytoskeleton. Localized in cytoplasmic mRNP granules containing untranslated mRNAs.

Target information above from: UniProt accession P60709 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
• Database links
  • Entrez Gene: 60 Human
  • Entrez Gene: 11461 Mouse
  • Entrez Gene: 100009272 Rabbit
  • Entrez Gene: 81822 Rat
  • Entrez Gene: 398459 Xenopus laevis
  • Omim: 102630 Human
  • SwissProt: P60706 Chicken
  • SwissProt: P60712 Cow

  • SwissProt: P60709 Human
  • SwissProt: P60710 Mouse
  • SwissProt: P29751 Rabbit
  • SwissProt: P60711 Rat
  • SwissProt: O93400 Xenopus laevis
  • Unigene: 520640 Human
  • Unigene: 708120 Human
  • Unigene: 727576 Human
  • Unigene: 328431 Mouse
  • Unigene: 391967 Mouse
  • Unigene: 94978 Rat
  • Unigene: 4138 Xenopus laevis

  see all

• Alternative names
  A X actin like protein antibodyACTB antibodyACTB_HUMAN antibody

  Actin antibodyActin cytoplasmic 1 antibodyActx antibodyb-actin antibodyBeta cytoskeletal actin antibodyBeta-actin antibodycytoplasmic 1 antibodyMelanoma X actin antibodyN-terminally processed antibodyPS1TP5 binding protein 1 antibodyPS1TP5BP1 antibody

  see all

Anti-beta Actin antibody [mAbcam 8224] - Loading Control images

• 

  Western blot - beta Actin antibody [mAbcam 8224] - Loading Control (ab8224)
  All lanes : Anti-beta Actin antibody [mAbcam 8224] - Loading Control (ab8224)
  
  Lane 1 : Drosophila lysate at 20 µg
  Lane 2 : S. pombe lysate
  Lane 3 : S. cerevisiae lysate (Actin 1 - please see note)
  
  Secondary
  Rabbit polyclonal Secondary Antibody to Mouse IgG - H&L (HRP) (ab6728) at 1/5000 dilution
  developed using the ECL technique
  
  Performed under reducing conditions.
  
  Predicted band size : 42 kDa
  
  
  Note: although S. cerevisae is not known to express beta Actin, Abcam believes that the band on lane 3 corresponds to Actin 1 (Swissprot ID: P60010, based on sequence similarity).
• 

  Western blot - beta Actin antibody [mAbcam 8224] - Loading Control (ab8224)
  Anti-beta Actin antibody [mAbcam 8224] - Loading Control (ab8224) + Xenopus embryo lysate at 20 µg
  
  Secondary
  Rabbit polyclonal Secondary Antibody to Mouse IgG - H&L (HRP) (ab6728)
  developed using the ECL technique
  
  Performed under reducing conditions.
  
  Predicted band size : 42 kDa
  
• 

  Western blot - beta Actin antibody [mAbcam 8224] - Loading Control (ab8224)
  All lanes : Anti-beta Actin antibody [mAbcam 8224] - Loading Control (ab8224) at 1/1000 dilution
  
  Lane 1 : Fruit fly (Drosophila melanogastor) whole cell lysate - Female
  Lane 2 : Fruit fly (Drosophila melanogastor) whole cell lysate - Male
  
  Lysates/proteins at 100 µg per lane.
  
  Secondary
  An HRP-conjugated Sheep polyclonal to mouse IgG at 1/10000 dilution
  developed using the ECL technique
  
  Performed under reducing conditions.
  
  Predicted band size : 42 kDa
  
  
  Exposure time : 2 minutes

  This image is courtesy of an anonymous Abreview

  See Abreview

• 

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - beta Actin antibody [mAbcam 8224] - Loading Control (ab8224)Carl Hobbs, King`s College London, United Kingdom

  Immunohistochemistical detection of beta Actin using antibody [mAbcam 8224] - Loading Control on formaldehyde-fixed paraffin-embedded rat cerebellum sections. Antigen retrieval step: heat mediated Citric acid pH6 buffer.  Permeabilization: No. Blocking step: 1% BSA for 10 mins @ rt°C. Primary antibody dilution 1/1000 for 2 hours in TBS/BSA/azide. Secondary Antibody: anti Mouse Igs conjugated to biotin (1/200). beta Actin appears to be particularly enriched not only in the glomeruli of the Granule cell layer (indicated by red arrowheads ) but also in Microglia (indicated by green arrowheads); All positive microglia appear to be ramified thus not presumed to be activated.

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• 

  Immunocytochemistry/ Immunofluorescence - Anti-beta Actin antibody [mAbcam 8224] - Loading Control (ab8224)
  ICC/IF image of ab8224 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab8224, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
  
  
• 

  Flow Cytometry-Anti-beta Actin antibody [mAbcam 8224] - Loading Control(ab8224)
  Overlay histogram showing HeLa cells stained with ab8224 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab8224, 1µg/1x10⁶ cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was Mouse IgG3 [MG3-35] (ab18394, 1µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed.
  
  
• 

  Immunocytochemistry/ Immunofluorescence - Anti-beta Actin antibody [mAbcam 8224] - Loading Control (ab8224)This image is courtesy of an Abreview submitted by Armen Petrosyan
  Immunofluorescence analysis of Human Panc-1 cells, staining beta Actin with ab8224.
  
  Cells were fixed with formaldehyde and blocked with 1% donkey serum in PBST for 1 hour at 22°C. Samples were incubated with primary antibody (1/75 in 1% donkey serum in PBST) for 1 hour at 22°C. A DyLight®488-conjugated donkey anti-mouse polyclonal IgG (1/200) was used as the secondary antibody.

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