Product nameAnti-alpha Actinin 4 antibody [EPR2533(2)]
See all alpha Actinin 4 primary antibodies
DescriptionRabbit monoclonal [EPR2533(2)] to alpha Actinin 4
Tested applicationsSuitable for: Flow Cyt, WB, IP, IHC-P, ICC/IFmore details
Species reactivityReacts with: Mouse, Rat, Human
Synthetic peptide within Human alpha Actinin 4 aa 1-100 (N terminal). The exact sequence is proprietary.
- Fetal skeletal muscle, A431, MCF-7, and HeLa cell lysates; Human kidney tissue; HeLa cells ICC-IF: HAP1 cells (HAP1-ACTN4 knockout cells used as negative cell line)
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
Storage bufferpH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol, 0.05% BSA, 50% Tissue culture supernatant
PurityTissue culture supernatant
- Anti-alpha Actinin 4 antibody [EPR2533(2)] (Alexa Fluor® 488) (ab198608)
- Anti-alpha Actinin 4 antibody [EPR2533(2)] (Alexa Fluor® 647) (ab198610)
- Anti-alpha Actinin 4 antibody [EPR2533(2)] (HRP) (ab199072)
- Anti-alpha Actinin 4 antibody [EPR2533(2)] - BSA and Azide free (ab204919)
- Anti-alpha Actinin 4 antibody [EPR2533(2)] (Phycoerythrin) (ab223936)
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.
|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. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
|ICC/IF||1/100 - 1/250.|
FunctionF-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 specificityWidely expressed.
Involvement in diseaseDefects 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 similaritiesBelongs 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 localizationNucleus. 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
- actinin 4 antibody
- Actinin alpha 4 antibody
- actinin4 antibody
ab108198 staining ACTN4 in wild-type HAP1 cells (top panel) and ACTN4 knockout HAP1 cells (bottom panel). The cells were fixed with PFA (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab108198 at 1/100 dilution and ab7291 (Tubulin) at 1/1000 dilution overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
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.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ab108198, at 1/100 dilution, staining alpha Actinin 4 in HeLa cells by Immunofluorescence.
This product has been referenced in:
- Yin W et al. Protein kinase C and protein kinase A are involved in the protection of recombinant human glucagon-like peptide-1 on glomeruli and tubules in diabetic rats. J Diabetes Investig 10:613-625 (2019). Read more (PubMed: 30307132) »
- Schell C et al. ARP3 Controls the Podocyte Architecture at the Kidney Filtration Barrier. Dev Cell 47:741-757.e8 (2018). Read more (PubMed: 30503751) »