Anti-beta Actin antibody - Loading Control (ab8227)
Overview
- Product nameAnti-beta Actin antibody - Loading ControlSee all beta Actin primary antibodies ...
- DescriptionRabbit polyclonal to beta Actin - Loading Control
- Tested applicationsIHC-Fr, IP, WB, ICC, IHC-FrFl, IHC-P, ICC/IF, ELISA more details
- Species reactivityReacts with: Mouse, Rat, Sheep, Rabbit, Chicken, Guinea pig, Cow, Dog, Human, Pig, Xenopus laevis, Fish, Monkey, Rhesus monkey, Chinese Hamster
Does not react with
Zebrafish - Immunogen
Synthetic peptide derived from within residues 1 - 100 of Human beta Actin.
(Peptide available as ab13772.)
- Positive controlTested on HeLa whole cell lysate and mouse brain lysate
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: 1% BSA, PBS, pH 7.4 -
Concentration information loading... - PurityImmunogen affinity purified
- Clonality Polyclonal
- IsotypeIgG
- Research Areas
Applications
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 | Notes |
|---|---|
| IHC-Fr | IHC-Fr: Use at an assay dependent concentration. |
| IP | IP: Use at an assay dependent concentration. |
| WB | WB: 1/1000 - 1/5000. Can be blocked with beta Actin peptide (ab13772). |
| ICC | ICC: Use at an assay dependent concentration. |
| IHC-FrFl | IHC-FrFl: Use at an assay dependent concentration. |
| IHC-P | IHC-P: Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
| ICC/IF | ICC/IF: Use a concentration of 1 µg/ml. |
| ELISA | ELISA: 1/1000. |
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.
-
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: P60713 Sheep
- 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
Target information above from: UniProt accession
P60709
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010)
.
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Alternative names
- A X actin like protein antibodyACTB antibodyACTB_HUMAN antibody
- Actin antibodyActin cytoplasmic 1 antibodyActx antibodyb actin 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 - Loading Control images
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Western blot - beta Actin antibody - Loading Control (ab8227)All lanes : Anti-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 Secondary Antibody 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?) -
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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Western blot - beta Actin antibody - Loading Control (ab8227)All lanes : Anti-beta Actin antibody - Loading Control (ab8227) at 1/1000 dilution
Lane 1 : HeLa nuclear lysate
Lane 2 : HeLa whole cell lysate
Lane 3 : A431 cell lysate
Lane 4 : Jurkat cell lysate
Lane 5 : HEK 293 cell lysate
Lane 6 : HeLa nuclear lysate withbeta Actin peptide (ab13772) at 1 µg/ml
Lane 7 : HeLa whole cell lysate withbeta Actin peptide (ab13772) at 1 µg/ml
Lane 8 : A431 cell lysate withbeta Actin peptide (ab13772) at 1 µg/ml
Lane 9 : Jurkate cell lysate withbeta Actin peptide (ab13772) at 1 µg/ml
Lane 10 : HEK 293 cell lysate withbeta Actin peptide (ab13772) at 1 µg/ml
Lysates/proteins at 20 µg per lane.
Secondary
Goat polyclonal Secondary Antibody to 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 -
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. -
Immunohistochemistry (Frozen sections) - beta Actin antibody - Loading Control (ab8227)This image is courtesy of an anonymous abreview.ab8227 staining beta Actin in human stomach tissue sections by Immunohistochemistry (frozen sections). Tissue was fixed with acetone and then blocked with 5% serum for 1 hour at 23°C followed by incubation with the primary antibody, at 1µg/ml, for 1 hour at 23°C. An undiluted HRP-conjugated goat polyclonal was used as secondary antibody. -
ELISA - beta Actin antibody - Loading Control (ab8227)Image courtesy of an anonymous Abreview.ab8227 used in Direct ELISA in NIH 3T3 murine fibroblasts. Primary antibody used at a 1/1000 dilution for 16 hours at 4°C. The secondary antibody used is an AP-conjugated Goat anti-rabbit used at a 1/2000 dilution. A blocking step was performed using 5% BSA for 1 hour at 23°C. -
Immunocytochemistry/ Immunofluorescence - beta Actin antibody - Loading Control (ab8227)ICC/IF image of ab8227 stained HeLa cells. The cells were 100% methanol fixed (5 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 (ab8227, 5µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-rabbit IgG - H&L, pre-adsorbed (ab96899) used at a 1/250 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. -
Western blot - beta Actin antibody - Loading Control (ab8227)All lanes : Anti-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
Goat polyclonal Secondary Antibody to 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
References for Anti-beta Actin antibody - Loading Control (ab8227)
This product has been referenced in:
- Johannessen TC et al. The DNA repair protein ALKBH2 mediates temozolomide resistance in human glioblastoma cells. Neuro Oncol 15:269-78 (2013). Read more (PubMed: 23258843) »
- Schulz R et al. Inhibiting the HSP90 chaperone destabilizes macrophage migration inhibitory factor and thereby inhibits breast tumor progression. J Exp Med 209:275-89 (2012). Read more (PubMed: 22271573) »
