Anti-alpha Actinin 4 抗体 [7H6] (ab32816)

製品の概要

  • 製品名Anti-alpha Actinin 4 antibody [7H6]
    alpha Actinin 4 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [7H6] to alpha Actinin 4
  • アプリケーション適用あり: ELISA, Flow Cyt, IHC-P, WB, ICC/IFmore details
  • 種交差性
    交差種: Mouse, Human
    交差が予測される動物種: Rat, Cow, Dog, Pig, Xenopus laevis
  • 免疫原

    Synthetic peptide:

    APYQGPDAVPGALD

    , corresponding to amino acids 884-897 of Human alpha Actinin 4.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab32816 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
ELISA Use at an assay dependent concentration. PubMed: 19016781
Flow Cyt Use 1µg for 106 cells.
IHC-P Use a concentration of 4 µg/ml.
WB Use a concentration of 2 µg/ml. Predicted molecular weight: 105 kDa.
ICC/IF Use a concentration of 4 µg/ml.

ターゲット情報

  • 機能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.
  • 組織特異性Widely expressed.
  • 関連疾患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.
  • 配列類似性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.
  • 細胞内局在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
  • 参照データベース
  • 別名
    • 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

Anti-alpha Actinin 4 antibody [7H6] 画像

  • ab32816 (4 µg/ml) staining alpha actinin in human lung, using an automated system (DAKO Autostainer Plus). Using this protocol there is strong staining of both cytoplasm and nuclei of the bronchial epithelium and resident macrophage cells.
    Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
  • Overlay histogram showing HeLa cells stained with ab32816 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab32816, 1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was a goat anti-mouse DyLight® 488 (IgG; H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was a mix of mouse IgG1 [ICIGG1], (ab91353, 1μg/1x106 cells), IgG2a [ICIGG2A], (ab91361, 1μg/1x106 cells), IgG2b [PLPV219], (ab91366, 1μg/1x106 cells), IgG3 [MG3-35], (ab18394, 1μg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized Human colon carcinoma tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:200 with a mouse monoclonal antibody recognizing alpha Actinin 4 ab32816 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

  • Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized Human kidney tissue tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:200 with a mouse monoclonal antibody recognizing alpha Actinin 4 ab32816 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

  • Immunohistochemistry was performed on both normal and cancer biopsies of deparaffinized Human tonsil tissue tissues. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:200 with a mouse monoclonal antibody recognizing alpha Actinin 4 ab32816 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

  • Immunofluorescence analysis of LNCaP cells, staining alpha Actinin 4 with ab32816.

    Cells were fixed with formaldehyde and blocked with 1% serum for 1 hour at 22°C. Samples were incubated with primary antibody (1/100 in 1% donkey serum in PBST) for 1 hour at 22°C. An undiluted DyLight®488-conjugated donkey anti-mouse polyclonal IgG was used as the secondary antibody.

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Anti-alpha Actinin 4 antibody [7H6] (ab32816) 使用論文

This product has been referenced in:
  • Piskareva O  et al. The development of cisplatin resistance in neuroblastoma is accompanied by epithelial to mesenchymal transition in vitro. Cancer Lett N/A:N/A (2015). Read more (PubMed: 25960282) »
  • Lenhart KC  et al. GRAF1 deficiency blunts sarcolemmal injury repair and exacerbates cardiac and skeletal muscle pathology in dystrophin-deficient mice. Skelet Muscle 5:27 (2015). ICC/IF ; Mouse . Read more (PubMed: 26301073) »

See all 5 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (LNCaP)
Specification LNCaP
Fixative Formaldehyde
Permeabilization No
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 22°C
Username

Dr. Armen Petrosyan

Verified customer

投稿 Nov 14 2012

Unfortunately we have not determined the isotype of the Alpha Actinin 4 antibody (ab32816).

For ab98134 DISCOUNT CODE: ***
For ab32816 DISCOUNT CODE: ***
Expiration date: November 10th, 2012

I am very pleased to hear you would like to use ab98134 inWB and ab32816 in IFand send us an image of your results.

Each co...

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Thank you for your query regarding antibody ab32816. As far as I am aware, at this time, ab32816 has not tested cross-reactivity to actin 2, therefore there is no data regarding whether or not the product will or will not cross react with actinin 2. U...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"