Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-alpha Actinin 4 antibody [EPR2533(2)] - BSA and Azide free (ab204919)

Overview

  • Product name

    Anti-alpha Actinin 4 antibody [EPR2533(2)] - BSA and Azide free
    See all alpha Actinin 4 primary antibodies
  • Description

    Rabbit monoclonal [EPR2533(2)] to alpha Actinin 4 - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, WB, IP, Flow Cyt, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human alpha Actinin 4. The exact sequence is proprietary.

  • General notes

    Ab204919 is the carrier-free version of ab108198. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab204919 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

Properties

Applications

Our Abpromise guarantee covers the use of ab204919 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
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB Use at an assay dependent concentration. Detects a band of approximately 100 kDa (predicted molecular weight: 105 kDa).
IP Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG (Low endotoxin, Azide free), is suitable for use as an isotype control with this antibody.

 

ICC/IF Use at an assay dependent concentration.

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

  • This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108198).

    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)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108198).

  • ab108198, at 1/100 dilution, staining alpha Actinin 4 in HeLa cells by Immunofluorescence.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108198).

  • This IHC data was generated using the same anti-alpha Actinin 4 antibody clone, EPR2533(2), in a different buffer formulation (cat# ab108198).

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

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

References

ab204919 has not yet been referenced specifically in any publications.

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