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リコンビナントRabMAb

Anti-NOTCH3 抗体 [EPR25183-101] (ab300527)

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Western blot - Anti-NOTCH3 antibody [EPR25183-101] (AB300527)
  • Western blot - Anti-NOTCH3 antibody [EPR25183-101] (AB300527)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NOTCH3 antibody [EPR25183-101] (AB300527)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NOTCH3 antibody [EPR25183-101] (AB300527)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NOTCH3 antibody [EPR25183-101] (AB300527)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NOTCH3 antibody [EPR25183-101] (AB300527)
  • Flow Cytometry (Intracellular) - Anti-NOTCH3 antibody [EPR25183-101] (AB300527)
  • Anti-NOTCH3 antibody [EPR25183-101] (AB300527)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR25183-101] to NOTCH3
  • Suitable for: WB, IHC-P, Flow Cyt (Intra)
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

製品の概要

  • 製品名

    Anti-NOTCH3 antibody [EPR25183-101]
    NOTCH3 一次抗体 製品一覧

  • 製品の詳細

    Rabbit monoclonal [EPR25183-101] to NOTCH3

  • 由来種

    Rabbit

  • アプリケーション

    適用あり: WB, IHC-P, Flow Cyt (Intra)more details

  • 種交差性

    交差種: Human
    非交差種: Mouse, Rat

  • 免疫原

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • ポジティブ･コントロール

    • WB: HeLa transfected with scrambled siRNA control; MCF7 whole cell lysate. IHC-P: Human endometrium; human smooth muscle; human lung carcinoma tissue. HEK-293T cells transfected with a human NOTCH3 expression vector containing a his tag in human tissue. Flow Cyt (Intra): MCF7 cells.

  • 特記事項

    AB300527 does not react in WB with mouse, IHC-P with mouse and rat, ICC with mouse, and intracellular flow cytometry with mouse. 

    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.

製品の特性

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab300527の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
WB (1)
1/1000. Predicted molecular weight: 244 kDa.
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cyt (Intra)
1/50.
特記事項
WB
1/1000. Predicted molecular weight: 244 kDa.
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cyt (Intra)
1/50.

ターゲット情報

  • 機能

    Functions as a receptor for membrane-bound ligands Jagged1, Jagged2 and Delta1 to regulate cell-fate determination. Upon ligand activation through the released notch intracellular domain (NICD) it forms a transcriptional activator complex with RBPJ/RBPSUH and activates genes of the enhancer of split locus. Affects the implementation of differentiation, proliferation and apoptotic programs.

  • 組織特異性

    Ubiquitously expressed in fetal and adult tissues.

  • 関連疾患

    Defects in NOTCH3 are the cause of cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) [MIM:125310]. CADASIL causes a type of stroke and dementia of which key features include recurrent subcortical ischemic events and vascular dementia. The disorder affects relatively young adults of both sexes. Mutations affect highly conserved cysteine residues within epidermal growth factor (EGF)-like repeat domains in the extracellular part of the receptor.

  • 配列類似性

    Belongs to the NOTCH family.
    Contains 5 ANK repeats.
    Contains 34 EGF-like domains.
    Contains 3 LNR (Lin/Notch) repeats.

  • 翻訳後修飾

    Synthesized in the endoplasmic reticulum as an inactive form which is proteolytically cleaved by a furin-like convertase in the trans-Golgi network before it reaches the plasma membrane to yield an active, ligand-accessible form. Cleavage results in a C-terminal fragment N(TM) and a N-terminal fragment N(EC). Following ligand binding, it is cleaved by TNF-alpha converting enzyme (TACE) to yield a membrane-associated intermediate fragment called notch extracellular truncation (NEXT). This fragment is then cleaved by presenilin dependent gamma-secretase to release a notch-derived peptide containing the intracellular domain (NICD) from the membrane.
    Phosphorylated.

  • 細胞内局在

    Cell membrane and Nucleus. Following proteolytical processing NICD is translocated to the nucleus.
  • Information by UniProt

  • 参照データベース

    • 別名

      • CADASIL antibody
      • CASIL antibody
      • NOTC3_HUMAN antibody
      • Notch 3 antibody
      • Notch 3 intracellular domain antibody
      • Notch homolog 3 antibody
      • Notch3 antibody
      see all

    画像

    • Western blot - Anti-NOTCH3 antibody [EPR25183-101] (AB300527)
      Western blot - Anti-NOTCH3 antibody [EPR25183-101] (AB300527)
      All lanes : Anti-NOTCH3 antibody [EPR25183-101] (ab300527) at 1/1000 dilution

      Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) transfected with scrambled siRNA control, whole cell lysate
      Lane 2 : HeLa transfected with siRNA specifically targeting NOTCH3, whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Developed using the ECL technique.

      Predicted band size: 244 kDa
      Observed band size: 90, 270 kDa why is the actual band size different from the predicted?


      Exposure time: 169 seconds


      Blocking / Diluting buffer: 5% NFDM/TBST.

      The antibody recognises full-length (FL) Notch3 at 270KDa and a membrane tethered Notch3 fragment (NTM) at 90 KDa.

      This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

    • Western blot - Anti-NOTCH3 antibody [EPR25183-101] (AB300527)
      Western blot - Anti-NOTCH3 antibody [EPR25183-101] (AB300527)
      All lanes : Anti-NOTCH3 antibody [EPR25183-101] (ab300527) at 1/1000 dilution

      Lane 1 : MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate at 26 µg
      Lane 2 : MDA-MB-231 (human breast adenocarcinoma epithelial cell), whole cell lysate at 20 µg

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Developed using the ECL technique.

      Predicted band size: 244 kDa
      Observed band size: 90, 270 kDa why is the actual band size different from the predicted?


      Exposure time: 48 seconds


      Blocking / Diluting buffer: 5% NFDM/TBST.

      Low expression: MDA-MB-231 (PMID: 29109797).

      The antibody recognises full-length (FL) Notch3 at 270KDa and a membrane tethered Notch3 fragment (NTM) at 90 KDa.

      The expression profile and molecular weight observed are consistent with what has been described in the literature (PMID: 29109797, 24743243, 25356737).

      Lysates were freshly made and used immediately to minimize protein degradation.

      This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NOTCH3 antibody [EPR25183-101] (AB300527)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NOTCH3 antibody [EPR25183-101] (AB300527)

      Immunohistochemical analysis of paraffin-embedded human endometrium tissue labeling NOTCH3 with ab300527 at 1/500 dilution (1.012 µg/ml) followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection) Kit. Positive staining on vascular smooth muscle cells of human endometrium. The section was incubated with ab300527 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NOTCH3 antibody [EPR25183-101] (AB300527)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NOTCH3 antibody [EPR25183-101] (AB300527)

      Immunohistochemical analysis of paraffin-embedded human smooth muscle tissue labeling NOTCH3 with ab300527 at 1/500 dilution (1.012 µg/ml) followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection) Kit. positive staining on vascular smooth muscle cells in human smooth muscle. The section was incubated with ab300527 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NOTCH3 antibody [EPR25183-101] (AB300527)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NOTCH3 antibody [EPR25183-101] (AB300527)

      Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue labeling NOTCH3 with ab300527 at 1/500 dilution (1.012 µg/ml) followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection) Kit. Positive staining on Human lung carcinoma. The section was incubated with ab300527 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NOTCH3 antibody [EPR25183-101] (AB300527)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NOTCH3 antibody [EPR25183-101] (AB300527)

      Immunohistochemical analysis of NOTCH3 using paraffin-embedded human tissue.

      Panel A: HEK-293T cells transfected with a human NOTCH3 expression vector containing a his tag.

      Panel B: HEK-293T cells transfected with empty vector containing a his tag.

      NOTCH3 labeled with ab300527 at 1/1000 dilution (0.506 µg/ml) followed by a ready to use Leica DS9800 (BOND™ Polymer Refine Detection) Kit. Positive staining on (A) HEK-293T transfected with a human NOTCH3 expression vector, no staining on (B) HEK-293T transfected with empty vector. The section was incubated with ab300527 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (BOND™ Polymer Refine Detection).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

    • Flow Cytometry (Intracellular) - Anti-NOTCH3 antibody [EPR25183-101] (AB300527)
      Flow Cytometry (Intracellular) - Anti-NOTCH3 antibody [EPR25183-101] (AB300527)

      Flow Cytometry (Intracellular) analysis of MDA-MB-231 (human breast adenocarcinoma epithelial cells (Left)) and MCF7 (human breast adenocarcinoma epithelial cells (Right)) labeling NOTCH3 (red) with ab300527 at a 1/50 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A Goat Anti-Rabbit IgG (Alexa Fluor® 488, (ab150081)) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730). Blue - (unlabeled control) - Cells without incubation with primary and secondary antibodies. 

      Low expression: MDA-MB-231(PMID: 29109797).

    • Anti-NOTCH3 antibody [EPR25183-101] (AB300527)
      Anti-NOTCH3 antibody [EPR25183-101] (AB300527)

    プロトコール

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    参考文献 (0)

    ab300527 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

    ab300527 は論文での使用が確認できていません。

    レビューと Q&A

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    レビューを送る 質問を送る

    Western blot abreview for Anti-NOTCH3 antibody [EPR25183-101]

     Excellent
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (hiPSCs)
    Gel Running Conditions
    Non-reduced Denaturing (4-15%)
    Loading amount
    20 µg
    Specification
    hiPSCs
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    MS. Igor Neves Barbosa

    Verified customer

    投稿 Oct 25 2022

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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