重组Anti-alpha Actinin 4抗体[EPR2533(2)] (ab108198)
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: Human
概述
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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 -
宿主
Rabbit -
Tested Applications & Species
Application Species Flow Cyt HumanICC/IF HumanIHC-P HumanIP HumanWB Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- 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
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常规说明
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.
性能
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形式
Liquid -
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. -
存储溶液
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant -
Concentration information loading...
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纯度
Tissue culture supernatant -
克隆
单克隆 -
克隆编号
EPR2533(2) -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
- Alexa Fluor® 488 Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab198608)
- Alexa Fluor® 647 Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab198610)
- HRP Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab199072)
- Anti-alpha Actinin 4 antibody [EPR2533(2)] - BSA and Azide free (ab204919)
- PE Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab223936)
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
应用
The Abpromise guarantee
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.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
应用 | Species |
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Flow Cyt |
Human
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ICC/IF |
Human
|
IHC-P |
Human
|
IP |
Human
|
WB |
Human
|
All applications |
Mouse
Rat
|
应用 | Ab评论 | 说明 |
---|---|---|
Flow Cyt |
Use at an assay dependent concentration.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
|
WB | (4) |
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 | (3) |
1/100 - 1/250.
|
说明 |
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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. |
靶标
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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. -
组织特异性
Widely expressed. -
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
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数据库链接
- Entrez Gene: 81 Human
- Entrez Gene: 60595 Mouse
- Entrez Gene: 63836 Rat
- Omim: 604638 Human
- SwissProt: O43707 Human
- SwissProt: P57780 Mouse
- SwissProt: Q9QXQ0 Rat
- Unigene: 270291 Human
see all -
别名
- actinin 4 antibody
- Actinin alpha 4 antibody
- actinin4 antibody
see all
图片
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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).
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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.
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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)
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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 -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha Actinin 4 antibody [EPR2533(2)] (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.
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ab108198, at 1/100 dilution, staining alpha Actinin 4 in HeLa cells by Immunofluorescence.
实验方案
数据表及文件
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SDS download
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Datasheet download
文献 (12)
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