Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR2533(2)] to alpha Actinin 4
- Suitable for: Flow Cyt, WB, IP, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
产品名称Anti-alpha Actinin 4抗体[EPR2533(2)]
参阅全部 alpha Actinin 4 一抗
描述兔单克隆抗体[EPR2533(2)] to alpha Actinin 4
经测试应用适用于: Flow Cyt, WB, IP, IHC-P, ICC/IFmore details
种属反应性与反应: 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) IP: HeLa cell lysate
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
存放说明Shipped 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.
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant
Concentration information loading...
纯度Tissue 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)] (PE) (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.|
功能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.
疾病相关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.
序列相似性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.
细胞定位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
- 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)
Purified ab108198 at 1/50 dilution (2µg) immunoprecipitating alpha Actinin 4 in HeLa whole cell lysate.
Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10µg
Lane 2 (+): ab108198 + HeLa whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab108198 in HeLa whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 105 kDa
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.
ab108198 被引用在 12 文献中.
- Burton KM et al. Dynamin 2 interacts with a-actinin 4 to drive tumor cell invasion. Mol Biol Cell 31:439-451 (2020). PubMed: 31967944
- Ko SY et al. Cancer-derived small extracellular vesicles promote angiogenesis by heparin-bound, bevacizumab-insensitive VEGF, independent of vesicle uptake. Commun Biol 2:386 (2019). PubMed: 31646189
- Dhanda AS et al. Listeria monocytogenes hijacks CD147 to ensure proper membrane protrusion formation and efficient bacterial dissemination. Cell Mol Life Sci 76:4165-4178 (2019). PubMed: 31076805
- 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). PubMed: 30307132
- Schell C et al. ARP3 Controls the Podocyte Architecture at the Kidney Filtration Barrier. Dev Cell 47:741-757.e8 (2018). PubMed: 30503751