使用敲除细胞株进行验证重组RabMAb

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

Reviews (6) Submit a question References (12)

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

概述

  • 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

  • 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

  • 免疫原

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

  • 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

  • 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
    For more information see here.

    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.

性能

应用

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.

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 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

  • 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

  • 数据库链接

    see all

  • 别名

    • 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

图片

  • Immunocytochemistry/ Immunofluorescence - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198)
    Immunocytochemistry/ Immunofluorescence - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198)

    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).

  • Western blot - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198)
    Western blot - Anti-alpha Actinin 4 antibody [EPR2533(2)] (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.

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

    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)

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

    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

  • Western blot - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198)
    Western blot - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198)
    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)
    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.

  • Immunocytochemistry/ Immunofluorescence - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198)
    Immunocytochemistry/ Immunofluorescence - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198)
    ab108198, at 1/100 dilution, staining alpha Actinin 4 in HeLa cells by Immunofluorescence.
  • Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198)
    Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198)

文献 (12)

发表研究结果有使用 ab108198？请让我们知道，以便我们可以引用本数据表中的参考文章。

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
View all Publications for this product

客户评价及客户问答

Show All 评价 Q&A
提交评价 提交问题

1-6 of 6 Abreviews or Q&A

Immunohistochemistry (Frozen sections) abreview for Anti-alpha Actinin 4 antibody [EPR2533(2)]

 Inconclusive
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Hippocampus (Neurons in the CA1 Region))
Permeabilization
Yes - 0,1% Triton in PBS
Specification
Hippocampus (Neurons in the CA1 Region)
Blocking step
Serum as blocking agent for 45 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Paraformaldehyde
Read More

Helena Lichtenfeld

Verified customer

提交于 Aug 21 2020

Immunocytochemistry abreview for Anti-alpha Actinin 4 antibody [EPR2533(2)]

 Excellent
Abreviews
abreview image
Application
Immunocytochemistry
Sample
Human Cell (HeLa cells)
Permeabilization
Yes - 0.5% Triton-X
Specification
HeLa cells
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 25°C
Fixative
Formaldehyde
Read More

Anugraha Rajagopalan

Verified customer

提交于 Oct 29 2018

Western blot abreview for Anti-alpha Actinin 4 antibody [EPR2533(2)]

 Excellent
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

提交于 Dec 02 2016

Western blot abreview for Anti-alpha Actinin 4 antibody [EPR2533(2)]

 Excellent
Abreviews
abreview image
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
Read More

Abcam user community

Verified customer

提交于 Nov 02 2015

Western blot abreview for Anti-alpha Actinin 4 antibody [EPR2533(2)]

 Excellent
Abreviews
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
Read More

Abcam user community

Verified customer

提交于 Aug 15 2012

Immunocytochemistry/ Immunofluorescence abreview for Anti-alpha Actinin 4 antibody [EPR2533(2)]

 Excellent
Abreviews
abreview image
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%
Read More

Abcam user community

Verified customer

提交于 Aug 15 2012

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com