Validated using a knockout cell line
Recombinant
RabMAb

Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198)

Overview

  • Product name
    Anti-alpha Actinin 4 antibody [EPR2533(2)]
    See all alpha Actinin 4 primary antibodies
  • Description
    Rabbit monoclonal [EPR2533(2)] to alpha Actinin 4
  • Host species
    Rabbit
  • Tested applications
    Suitable for: Flow Cyt, WB, IP, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human alpha Actinin 4 aa 1-100 (N terminal). The exact sequence is proprietary.

  • Positive control
    • Fetal skeletal muscle, A431, MCF-7, and HeLa cell lysates; Human kidney tissue; HeLa cells
  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab108198 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
Flow Cyt Use at an assay dependent concentration.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

WB 1/1000 - 1/10000. Detects a band of approximately 100 kDa (predicted molecular weight: 105 kDa).
IP 1/10 - 1/100.
IHC-P 1/100 - 1/250.
ICC/IF 1/100 - 1/250.

Target

  • Function
    F-actin cross-linking protein which is thought to anchor actin to a variety of intracellular structures. This is a bundling protein. Probably involved in vesicular trafficking via its association with the CART complex. The CART complex is necessary for efficient transferrin receptor recycling but not for EGFR degradation.
  • Tissue specificity
    Widely expressed.
  • Involvement in disease
    Defects in ACTN4 are the cause of focal segmental glomerulosclerosis type 1 (FSGS1) [MIM:603278]. A renal pathology defined by the presence of segmental sclerosis in glomeruli and resulting in proteinuria, reduced glomerular filtration rate and edema. Renal insufficiency often progresses to end-stage renal disease, a highly morbid state requiring either dialysis therapy or kidney transplantation.
  • Sequence similarities
    Belongs to the alpha-actinin family.
    Contains 1 actin-binding domain.
    Contains 2 CH (calponin-homology) domains.
    Contains 2 EF-hand domains.
    Contains 4 spectrin repeats.
  • Cellular localization
    Nucleus. Cytoplasm. Localized in cytoplasmic mRNP granules containing untranslated mRNAs. Colocalizes with actin stress fibers. Nuclear translocation can be induced by the PI3 kinase inhibitor wortmannin or by cytochalasin D. Exclusively localized in the nucleus in a limited number of cell lines.
  • Information by UniProt
  • Database links
  • Alternative names
    • actinin 4 antibody
    • Actinin alpha 4 antibody
    • actinin4 antibody
    • ACTN 4 antibody
    • ACTN4 antibody
    • ACTN4_HUMAN antibody
    • Alpha-actinin-4 antibody
    • DKFZp686K23158 antibody
    • F actin cross linking protein antibody
    • F-actin cross-linking protein antibody
    • Focal segmental glomerulosclerosis 1 antibody
    • FSGS 1 antibody
    • FSGS antibody
    • FSGS1 antibody
    • Non muscle alpha actinin 4 antibody
    • Non-muscle alpha-actinin 4 antibody
    see all

Images

  • Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
    Lane 2: ACTN4 (alpha Actinin 4) knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HeLa whole cell lysate (20 µg)
    Lane 4: MCF7 whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab108198 observed at 105 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab108198 was shown to specifically react with alpha Actinin 4 in wild-type HAP1 cells as signal was lost in ACTN4 (alpha Actinin 4) knockout cells. Wild-type and ACTN4 (alpha Actinin 4) knockout samples were subjected to SDS-PAGE. ab108198 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • ab108198 staining alpha Actinin 4 in the human cell line HeLa (human cervix adenocarcinoma) by flow cytometry. Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. The sample was incubated with the primary antibody at a dilution of 1/50. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody.

    Isoytype control: Rabbit monoclonal IgG (Black)

    Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)

  • All lanes : Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198) at 1/1000 dilution

    Lane 1 : Fetal skeletal muscle lysate
    Lane 2 : A431 cell lysate
    Lane 3 : MCF-7 cell lysate
    Lane 4 : HeLa cell lysate

    Lysates/proteins at 10 µg per lane.

    Predicted band size: 105 kDa

  • ab108198, at 1/100 dilution, staining alpha Actinin 4 in paraffin-embedded Human kidney tissue by Immunohistochemistry.
  • ab108198, at 1/100 dilution, staining alpha Actinin 4 in HeLa cells by Immunofluorescence.

References

This product has been referenced in:
  • Wahlund CJE  et al. Exosomes from antigen-pulsed dendritic cells induce stronger antigen-specific immune responses than microvesicles in vivo. Sci Rep 7:17095 (2017). WB ; Mouse . Read more (PubMed: 29213052) »
  • Hamill KJ  et al. Alpha actinin-1 regulates cell-matrix adhesion organization in keratinocytes: consequences for skin cell motility. J Invest Dermatol 135:1043-52 (2015). Read more (PubMed: 25431851) »

See all 4 Publications for this product

Customer reviews and Q&As

Application
Western blot
Sample
Human Cell lysate - whole cell (Vascular smooth muscle cells)
Gel Running Conditions
Reduced Denaturing
Loading amount
25 µg
Specification
Vascular smooth muscle cells
Blocking step
Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
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Verified customer

Submitted Dec 02 2016

Application
Western blot
Sample
Rat Cell lysate - other (Primary lung epithelia cells)
Gel Running Conditions
Reduced Denaturing (4-12% Nupage)
Specification
Primary lung epithelia cells
Blocking step
(agent) for 2 hour(s) and 0 minute(s) · Concentration: 0.2% · Temperature: 22°C
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Verified customer

Submitted Nov 02 2015

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Breast)
Loading amount
20 µg
Specification
Breast
Gel Running Conditions
Reduced Denaturing (10%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Abcam user community

Verified customer

Submitted Aug 15 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Breast)
Specification
Breast
Fixative
Formaldehyde
Permeabilization
Yes - 0.2% Triton X-100
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
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Verified customer

Submitted Aug 15 2012

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

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