Anti-Actin antibody - Loading Control (ab1801)
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- References (148)
- Protocols
Overview
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Product nameAnti-Actin antibody - Loading Control
See all Actin primary antibodies -
DescriptionRabbit polyclonal to Actin - Loading Control
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Host speciesRabbit
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SpecificityThis antibody recognises beta and gamma actin in Human samples. It probably also recognises all the other known forms of Human actin. This antibody detects a single clean band in Human, Mouse, Rat, Chicken and Drosophila samples. In Xenopus laevis a secondary band is detected at about 30kDa. We are unsure whether this is cross-reaction with another actin isoform or merely non-specific. In Cow a doublet is detected, which probably represents different forms of actin.
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Tested applicationsSuitable for: WB, IHC-Pmore details
Unsuitable for: ICC/IF -
Species reactivityReacts with: Mouse, Rat, Chicken, Cow, Human, Xenopus laevis, Drosophila melanogaster, Zebrafish
Predicted to work with: Rabbit, Saccharomyces cerevisiae, Orangutan
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Immunogen
Synthetic peptide conjugated to KLH derived from within residues 350 to the C-terminus of Human Actin.
Read Abcam's proprietary immunogen policy (Peptide available as ab13771.) -
Positive control
- HeLa whole cell lysate or mouse brain lysate. IHC-P - Human Colon FFPE tissue section
Properties
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FormLiquid
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Storage instructionsShipped 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.
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Storage bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 -
Concentration information loading... -
PurityImmunogen affinity purified
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ClonalityPolyclonal
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IsotypeIgG
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Research areas
Associated products
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
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Related Products
Applications
Our Abpromise guarantee covers the use of ab1801 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 |
|---|---|---|
| WB | 1/1000. Detects a band of approximately 42 kDa (predicted molecular weight: 42 kDa). Block in 5% BSA. Blocking in milk significantly reduces the signal. | |
| IHC-P | Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Target
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FunctionActins are highly conserved proteins that are involved in various types of cell motility and are ubiquitously expressed in all eukaryotic cells.
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Involvement in diseaseDefects in ACTA1 are the cause of nemaline myopathy type 3 (NEM3) [MIM:161800]. A form of nemaline myopathy. Nemaline myopathies are muscular disorders characterized by muscle weakness of varying severity and onset, and abnormal thread-or rod-like structures in muscle fibers on histologic examination. The phenotype at histological level is variable. Some patients present areas devoid of oxidative activity containg (cores) within myofibers. Core lesions are unstructured and poorly circumscribed.
Defects in ACTA1 are a cause of myopathy congenital with excess of thin myofilaments (MPCETM) [MIM:161800]. A congenital muscular disorder characterized at histological level by areas of sarcoplasm devoid of normal myofibrils and mitochondria, and replaced with dense masses of thin filaments. Central cores, rods, ragged red fibers, and necrosis are absent.
Defects in ACTA1 are a cause of congenital myopathy with fiber-type disproportion (CFTD) [MIM:255310]; also known as congenital fiber-type disproportion myopathy (CFTDM). CFTD is a genetically heterogeneous disorder in which there is relative hypotrophy of type 1 muscle fibers compared to type 2 fibers on skeletal muscle biopsy. However, these findings are not specific and can be found in many different myopathic and neuropathic conditions. -
Sequence similaritiesBelongs to the actin family.
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Cellular localizationCytoplasm > cytoskeleton.
- Information by UniProt
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Database links
- Entrez Gene: 421534 Chicken
- Entrez Gene: 281592 Cow
- Entrez Gene: 58 Human
- Entrez Gene: 11459 Mouse
- Entrez Gene: 29437 Rat
- Entrez Gene: 407658 Zebrafish
- Omim: 102610 Human
- SwissProt: P68139 Chicken
see all -
Alternative names
- a actin antibody
- ACTA antibody
- ACTA1 antibody
see all
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Actin antibody - Loading Control (ab1801)IHC image of ab1801 staining Actin in normal 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 ab1801, 5μl/ml 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 negative 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
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Western blot - Anti-Actin antibody - Loading Control (ab1801)All lanes : Anti-Actin antibody - Loading Control (ab1801) at 1 µg/ml
Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : Brain (Rat) Tissue Lysate
Lane 3 : Brain (Mouse) Tissue Lysate
Lane 4 :NIH 3T3 whole cell lysate (ab7179)
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 42 kDa
Observed band size: 42 kDa
Exposure time: 1 minuteThis 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 2% Bovine Serum Albumin before being incubated with ab1801 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406
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Western blot - Anti-Actin antibody - Loading Control (ab1801)Image from PLoS One. 2014; 9(3): e92128. Fig 9,•DOI: 10.1371/journal.pone.0092128Western blot analysis of HeLa cells treated for 12 hours with hesperidin (h) (2.5 μg/ml, 4,01 μM), mangiferin (5 μg/ml, 11.84 μM) (m), and hesperidin (2.5 μg/ml, 4.01 μM) in a presence of mangiferin (5 μg/ml, 11.84 μM) (h+m). Immunoblotting was performed with the following primary antibodies: Bax (ab32503), BCL2 (ab59348), beta actin (ab1801), and caspase 8. After the washing steps, the membranes were incubated with goat anti-rabbit IgG (H+L) or with goat anti-mouse IgG (H+L) HRP-conjugated secondary antibodies and detected using ECL. Densitometry was performed using Image Lab software v. 4.1 (BioRad).
Top panel: Following 12h of treatment of HeLa cells with hesperidin (h), mangiferin (m), and hesperidin in a presence of mangiferin (h+m), the mRNA levels were monitored in real - time PCR experiments. The BAX and BCL2 mRNA levels results from 2 independent experiments (n?=?8) are plotted relative to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and 18S rRNA levels and expressed as a fold change over the EtOH control. Error bars represent standard derivations.
Bottom panel: Following 12h of treatment of HeLa cells with hesperidin (h), mangiferin (m), and hesperidin in a presence of mangiferin (h+m), the protein levels of Bax and BCL2 were detected with SDS-PAGE and Western Blot and related to beta actin levels.
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Western blot - Anti-Actin antibody - Loading Control (ab1801)All lanes : Anti-Actin antibody - Loading Control (ab1801) at 1 µg/ml
Lane 1 : Brain (Mouse) Tissue Lysate (ab27253)
Lane 2 : NIH 3T3 (Mouse) Whole Cell Lysate (ab52956)
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 42 kDa
Exposure time: 30 seconds -
Western blot - Anti-Actin antibody - Loading Control (ab1801)This image is courtesy of an anonymous AbreviewAll lanes : Anti-Actin antibody - Loading Control (ab1801) at 1/1000 dilution
All lanes : Whole cell lysates prepared from HUVEC cells
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : HRP-conjugated goat polyclonal to rabbit Ig at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 42 kDa
Exposure time: 30 seconds
References
This product has been referenced in:
- Sánchez RG et al. Human and rodent temporal lobe epilepsy is characterized by changes in O-GlcNAc homeostasis that can be reversed to dampen epileptiform activity. Neurobiol Dis 124:531-543 (2019). Read more (PubMed: 30625365) »
- Marín M et al. Functional Comparison of XPF Missense Mutations Associated to Multiple DNA Repair Disorders. Genes (Basel) 10:N/A (2019). Read more (PubMed: 30658521) »
