Anti-beta Actin antibody - Loading Control (ab75186)

Overview

  • Product nameAnti-beta Actin antibody - Loading ControlSee all beta Actin primary antibodies ...
  • Description
    Rabbit polyclonal to beta Actin - Loading Control
  • Tested applicationsWB, IHC-P, ICC/IF more details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

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

    (Peptide available as ab13772.)

  • Positive control
    • This antibody gave a positive signal in the following whole cell lysates: Jurkat, A431, HEK293, HepG2, MCF-7, NIH3T3 and PC12. This antibody gave a positive result in IHC in the following FFPE tissue: Human normal skin and in HeLa cell line in IF.
  • General notesab75186 is raised using the same immunogen as ab8227. Batches sold under ab75186 have passed QC testing in Western blot only. In our hands, this product did not work in ICC/IF. For further information please contact our Customer Services team.

Properties

Applications

Our Abpromise guarantee covers the use of ab75186 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Notes
WB Use a concentration of 1 µg/ml. Detects a band of approximately 42 kDa (predicted molecular weight: 42 kDa).Can be blocked with Human beta Actin peptide (ab13772).
IHC-P Use a concentration of 5 µg/ml.
ICC/IF Use a concentration of 1 µg/ml.

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
    modifications
    ISGylated.
  • 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
  • Alternative names
    • A X actin like protein antibody
    • ACTB antibody
    • ACTB_HUMAN antibody
    • Actin antibody
    • Actin cytoplasmic 1 antibody
    • Actx antibody
    • b actin antibody
    • b-actin antibody
    • Beta cytoskeletal actin antibody
    • Beta-actin antibody
    • cytoplasmic 1 antibody
    • Melanoma X actin antibody
    • N-terminally processed antibody
    • PS1TP5 binding protein 1 antibody
    • PS1TP5BP1 antibody
    see all

Anti-beta Actin antibody - Loading Control images

  • All lanes : Anti-beta Actin antibody - Loading Control (ab75186) at 1 µg/ml

    Lane 1 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 2 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 3 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 4 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 5 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
    Lane 6 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate (ab7179)
    Lane 7 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution

    Performed under reducing conditions.

    Predicted band size : 42 kDa
    Observed band size : 42 kDa

  • developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 42 kDa


    Exposure time : 10 seconds

    Western blot image using the Optiblot Reducing Electrophoresis Kit - 10 x 10 cm (4-20%) (ab119220) with the
    Prism Ultra Protein Ladder (ab116028) 5µl used. We recommend using our ECL substrate kit (ab65623) .
    20ug of Lysate per lane and detection using ab75186 diluted 1/250.
    Lane 1: Hela cell lysate
    Lane 2: Jurkat cell lysate
    Lane 3: A431 cell lysate
    Lane 4: HEK293 cell lysate
    Lane 5: HepG2 cell lysate

  • IHC image of beta Actin staining in Human normal skin formalin fixed paraffin embedded tissue section, performed on a Leica 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 ab75186, 5µg/ml, 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.

    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.

  • ICC/IF image of ab75186 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 (ab75186, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) 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.

References for Anti-beta Actin antibody - Loading Control (ab75186)

This product has been referenced in:
  • Sasse SK  et al. The glucocorticoid receptor and KLF15 regulate gene expression dynamics and integrate signals through feed-forward circuitry. Mol Cell Biol 33:2104-15 (2013). WB . Read more (PubMed: 23508109) »
  • Calpe E  et al. ZAP-70 promotes the infiltration of malignant B-lymphocytes into the bone marrow by enhancing signaling and migration after CXCR4 stimulation. PLoS One 8:e81221 (2013). WB ; Human . Read more (PubMed: 24312539) »

See all 3 Publications for this product

Product Wall

Thank you for your reply.
We have beta actin antibodies that would fulfill all your criteria:
ab8227 (http://www.abcam.com/beta-Actin-antibody-Loading-Control-ab8227.html) - in case beta actin would be acceptable for your experiments.
Or...

Read More
Application Western blot
Sample Human Cell lysate - whole cell (HeLa and U2OS cells)
Loading amount 30 µg
Specification HeLa and U2OS cells
Gel Running Conditions Reduced Denaturing (12% Tris Glycine)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Verified customer

Submitted Jan 26 2011

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"